US20110099668A1

US20110099668A1 - Expressing GLK in plants

Info

Publication number: US20110099668A1
Application number: US12/151,046
Authority: US
Inventors: Jasbir Singh; Ghislaine C. Allard; Leonid V. Savitch; Rajagopal Subramaniam
Original assignee: Agriculture and Agri Food Canada AAFC
Current assignee: Agriculture and Agri Food Canada AAFC
Priority date: 2007-05-01
Filing date: 2008-05-01
Publication date: 2011-04-28
Also published as: CA2627079A1

Abstract

The present invention provides, in part, GLK1 nucleic acid molecules and polypeptides that can be used to confer resistance to a pathogen in a plant. The present invention also provides methods of detecting disease resistance genes and plants.

Description

FIELD OF INVENTION

The present invention relates to expressing GLK in plants. More specifically, the invention provides GLK1 molecules for conferring disease resistance to plants, increasing nitrogen status in plants, or both.

BACKGROUND OF THE INVENTION

The economic impact of plant pathogens in relation to agriculture and horticulture is considerable. For example, Fusarium Head Blight (FHB) is a major disease of concern for Canadian cereals.
Global gene expression analyses suggest that induced disease resistance can result from pathogen or pathogen derived molecule-induced reprogramming of gene networks (Li et al, 2006; Bae et al, 2006). The role of transcription factors in the modulation of these networks has been suggested for WRKY domain containing trans-factors (Li et al, 2006) and in the slh1 mutation (Naoutoshi et al, 2005).
Non-host resistance is the term used to describe the phenomenon where all members of a plant species exhibit resistance to all members of a given pathogen species. Non-host resistance is thought to be multigenic; therefore the inactivation of a single gene may not be sufficient to render a plant susceptible to a pathogen. Non-host resistance has been studied in salicylate hydroxylase (NahG) over-expressing plants, which convert salicylic acid to catechol and are defective in non-host response to pathogen. NahG plants expresses the salicylic acid degrading enzyme salicylate hydroxylase, resulting in indirect and side effects in plant defense (Heck et al, 2003; van Wees and Glazebrook 2003). Furthermore, it is known that part of non-host resistance is derived from “priming” of the plant to produce a readiness to infection.
The maize Golden2-like or GLK genes encode proteins belonging to a class of GARP domain of transcriptional activators (Hall, L. N. et al., 1998, Plant Cell 10:925-936; Fitter, W., et al., 2002, The Plant Journal 31: 713-727; Rossini, D. L., et al., 2001, Plant Cell 13: 1231-1244; US 2007/0022495) There are 56 predicted members of the GARP domain containing transcriptional activators in Arabidopsis (Riechmann, J. L., et al, 2000, Science 290: 2105-2110; which is incorporated herein by reference) and are involved in plant specific processes. These include the G2 or GLK proteins that are involved in chloroplast development (Cribb, L. et al., 2001, Genetics 159:787-797; Yasumura, Y., et al., 2005, Plant Cell 17: 1894-1907), the Arabidopsis type-B phosphorelay response regulators (ARR family) that mediate cytokinin signaling (Sakai, H., et al. Science 294:1519-1521; Hosoda, K., 2002, Plant Cell 14: 2015-2029; Mason, M. G., et al., 2005, Plant Cell 17:3007-3018), PSR1 proteins involved in phosphorus metabolism (Wykoff, D. D., et al., 1999, Proc. Natl. Acad. Sci. USA. 96:15336-15341), KANADI proteins in regulation of organ polarity (Kerstetter, R. A., et al., 2001, Nature 411:706-709) and b-ZIP G-box binding factors (Tamai, H., et al., 2002, Plant Cell Physiol. 43:99-107). Recently, increased accumulations of GLK1, 2 transcripts have been observed in constitutively frost tolerant transgenic Brassica napus overexpressing DREB1/CBFs, suggesting that it may play a role in temperature stress response and photosynthetic adaptation (Savitch, L. V., et al., 2005, Plant Cell Physiol. 46:1525-1539).

SUMMARY OF THE INVENTION

The present invention relates to expressing GLK in plants. More specifically, the invention provides GLK1 molecules for conferring disease resistance to plants, increasing nitrogen status in plants, or both.
The present invention provides a method of conferring resistance to a pathogen in a plant, the method comprising:
i) introducing a nucleic acid molecule encoding a GLK1 polypeptide or fragment or variant thereof into a plant cell to produce a transgenic plant cell, such that the nucleic acid molecule is positioned for expression in the transgenic plant cell; and
ii) growing a transgenic plant from the transgenic plant cell,
wherein the GLK1 polypeptide or fragment or variant thereof is over-expressed relative to wild type GLK1 at least 2 fold, whereby the 2 fold over-expression is sufficient to confer resistance to the pathogen in the plant.
The GLK1 may comprises the nucleic acid sequence set forth in SEQ ID NO: 1, a nucleic acid that exhibits an identity of from about 70 to about 100% with SEQ ID NO:1, or a nucleic acid molecule that hybridizes under conditions of high stringency to the nucleic acid sequence set forth in SEQ ID NO: 1. Furthermore, the GLK1 polypeptide may comprise an amino acid sequence substantially identical to the amino acid sequence set forth in SEQ ID NO: 2. Furthermore, the plant may be selected from the group consisting of canola, Brassica spp., maize, tobacco, alfalfa, potato, ginseng, pea, oat, rice, soybean, wheat, barley, sunflower, and cotton. The pathogen may be selected from the group consisting of Fusarium graminearum, Sclerotinia sclerotium, Puccinia graminis, Puccinia triticinia, Puccinia recondite, and Phytopthora infestans.
The present invention also provides a pathogen resistant plant comprising a GLK1 nucleic acid molecule that encodes a GLK1 polypeptide, wherein expression of the GLK1 produces at least 2 fold greater GLK1 polypeptide relative to a wild-type plant.
The present invention also provides a genetic construct comprising a regulatory region operably linked to a nucleic acid molecule encoding a GLK1 polypeptide or fragment or variant thereof. The nucleic acid molecule may comprise a nucleic acid sequence as set forth in SEQ ID NO: 1, a nucleotide sequence that exhibits from about 70% to about 100% sequence identity with the sequence of SEQ ID NO:1, or a nucleotide sequence that hybridizes to the nucleic acid sequence set forth in SEQ ID NO: 1 under conditions of high stringency. Furthermore, the GLK1 polypeptide may comprise an amino acid sequence set forth in SEQ ID NO: 2. Also provides is a vector comprising the genetic construct as just described. The vector may be pHS723-GLK1.
The present invention also pertains to a host cell, a transgenic plant, or a transgenic plant seed, comprising the genetic construct as described above. The host cell, transgenic plant, or a transgenic seed, may be selected from the group consisting of canola, Brassica spp., maize, tobacco, alfalfa, potato, ginseng, pea, oat, rice, soybean, wheat, barley, sunflower, and cotton. Furthermore, the transgenic plant is characterized as being resistant to a pathogen. The pathogen may be selected from the group consisting of Fusarium graminearum, Sclerotinia sclerotium, Puccinia graminis, Puccinia triticinia, Puccinia recondite, and Phytopthora infestans.
The present invention also provides a method of detecting a candidate pathogen-resistance gene in a plant, comprising, comparing the gene expression profile of a transgenic plant over-expressing a nucleic acid molecule encoding a GLK1 polypeptide or fragment or variant thereof, with the gene expression profile of a wild-type plant; and determining the genes over-expressed in the transgenic plant compared to the wild-type plant, wherein one or more than one of the genes over-expressed in the transgenic plant are candidate pathogen-resistance genes. The present invention also provides a method as just described wherein the GLK1 polypeptide is selected from the group consisting of an Arabidopsis GLK1, a wheat GLK1 polypeptide, a maize GLK1 polypeptide, a rice GLK1 polypeptide, and a soy bean GLK1 polypeptide.
The present invention also pertains to a method of detecting a plant exhibiting resistance to a pathogen, the method comprising:
i) determining the expression levels of a GLK1 nucleic acid molecule or polypeptide or fragment or variant thereof in a test plant;
ii) determining the expression levels of the GLK1 nucleic acid molecule or polypeptide or fragment or variant thereof in a control plant; and
iii) comparing the expression levels in the test plant to the expression levels in the control plant,
wherein the over-expression of the GLK1 nucleic acid molecule or polypeptide or fragment or variant thereof in the test plant compared to the control plant is indicative of the test plant being a plant exhibiting resistance to a pathogen. The test plant may be selected from the group consisting of Arabidopsis, canola, Brassica spp., maize, tobacco, alfalfa, potato, ginseng, pea, oat, rice, soybean, wheat, barley, sunflower, and cotton.
Furthermore the present invention includes a method of detecting a plant exhibiting resistance to a pathogen, the method comprising:
i) determining the expression levels of one or more of the nucleic acid molecule or polypeptide set out in Table 1 or in FIG. 3, or a fragment or variant thereof, in a test plant;
ii) determining the expression levels of the one or more of the nucleic acid molecule or polypeptide set out in Table 1 or in FIG. 3, or a fragment or variant thereof, in a control plant; and
iii) comparing the expression levels in the test plant to the expression levels in the control plant,
wherein the over-expression of the one or more of the nucleic acid molecule or polypeptide set out in Table 1 or in FIG. 3, or a fragment or variant thereof, in the test plant compared to the control plant is indicative of the test plant being a plant exhibiting resistance to a pathogen. The test plant is selected from the group consisting of Arabidopsis, canola, Brassica spp., maize, tobacco, alfalfa, potato, ginseng, pea, oat, rice, soybean, wheat, barley, sunflower, and cotton.
Also provides is a method of detecting a plant exhibiting resistance to a pathogen, the method comprising:
i) determining the expression levels of a At1 g77960, At2g41050, or both At1g77960, At2g41050, nucleic acid molecule or polypeptide or fragment or variant thereof in a test plant;
ii) determining the expression levels of the At1 g77960, At2g41050, or both At1g77960, At2g41050, nucleic acid molecule or polypeptide or fragment or variant thereof in a control plant; and
iii) comparing the expression levels in the test plant to the expression levels in the control plant,
wherein the over-expression of the At1g77960, At2g41050, or both At1g77960, At2g41050, nucleic acid molecule or polypeptide or fragment or variant thereof in the test plant compared to the control plant is indicative of the test plant being a plant exhibiting resistance to a pathogen. The test plant is selected from the group consisting of Arabidopsis, canola, Brassica spp., maize, tobacco, alfalfa, potato, ginseng, pea, oat, rice, soybean, wheat, barley, sunflower, and cotton.
The present invention also provides a method of increasing the nitrogen status in a plant, the method comprising:
i) introducing a nucleic acid molecule encoding a GLK1 polypeptide or fragment or variant thereof into a plant cell to produce a transgenic plant cell, such that the nucleic acid molecule is positioned for expression in the transgenic plant cell; and
ii) growing a transgenic plant from the transgenic plant cell,
wherein the GLK1 polypeptide or fragment or variant thereof is over-expressed relative to wild type GLK1 at least 2 fold, whereby the 2 fold over-expression is sufficient to confer resistance to the pathogen in the plant.
The GLK1 may comprises the nucleic acid sequence set forth in SEQ ID NO: 1, a nucleic acid that exhibits an identity of from about 70 to about 100% with SEQ ID NO:1, or a nucleic acid molecule that hybridizes under conditions of high stringency to the nucleic acid sequence set forth in SEQ ID NO: 1. Furthermore, the GLK1 polypeptide may comprise an amino acid sequence substantially identical to the amino acid sequence set forth in SEQ ID NO: 2. Furthermore, the plant may be selected from the group consisting of canola, Brassica spp., maize, tobacco, alfalfa, potato, ginseng, pea, oat, rice, soybean, wheat, barley, sunflower, and cotton.
The present invention also provides a plant with increased nitrogen status, the plant comprising a GLK1 nucleic acid molecule that encodes a GLK1 polypeptide, wherein expression of the GLK1 produces at least 2 fold greater GLK1 polypeptide relative to a wild-type plant.
The present invention also provides a plant, or a plant cell, that is characterized as having increase nitrogen status. The plant with increased nitrogen status comprising a GLK1 nucleic acid molecule that encodes a GLK1 polypeptide, wherein expression of the GLK1 produces at least 2 fold greater GLK1 polypeptide relative to a wild-type plant. The increase in nitrogen status determined by comparing the plant comprising the GLK1 nucleic acid molecule with a plant that does not express the GLK1 nucleic acid molecule and determining the levels of expression of ASN1, the colour of the leaves, or both determining the levels of expression of ASN1 and the colour of the leaves. Wherein an increase in the expression of ASN1, a darker green leaf, or both determining the levels of expression of ASN1 and the colour of the leaves, when compared to the plant that does not express GLK1, indicates an increase in nitrogen status.
The present invention provides a microarray comprising one or more of the nucleic acid molecule or polypeptide set out in Table 1 or in FIG. 3, or a fragment or variant thereof.
The target genes of GLK over-expression are described herein. The “regulon” of GLK1 was determined by over-expression of Arabidopsis GLK1 (AtGLK1) in Arabidopsis to encode and highly overexpress pathogen and pest related defense proteins (Table 1, FIG. 4). Arabidopsis AtGLK1 OE plants showed a phenotype with enhanced resistance to infection by the pathogen Fusarium graminearum (FIG. 3).
Affymetrix microarray analyses indicated that over-expression of wheat GLK1 (TaGLK1 Accession No. ABL10089) in Arabidopsis shows the regulation of similar genes as in AtGLK1 OE in Arabidopsis. These analyses indicate that TaGLK1 OE is capable of reprogramming identical networks of gene expression as that observed with AtGLK1 OE.
Transgenic wheat (Triticum aestivum cv Fielder) over-expressing wheat TaGLK1 also showed enhanced resistance to the spread of lesions in wheat leaves inoculated with F. graminearum and enhanced resistance in the development of Fusarium Head Blight (FHB) symptoms in developing wheat heads.
As described herein, overexpression of AtGLK1 and TaGLK1 in Arabidopsis also resulted in the upregulation of the glutamine dependent asparagine synthetase1 ASN1 transcripts. The over-expression of AtGLK1 in maize cv. Hii2 produced transgenic maize that had darker green leaves when grown under midwinter (early December, Latitude 46N) greenhouse conditions suggestive of enhanced nitrogen utilization. Over-expression of AtGLK1, TaGLK1,1 or a combination thereof, presents an opportunity to modify nitrogen status in crop plants.
This summary of the invention does not necessarily describe all features of the invention.

BRIEF DESCRIPTION OF THE DRAWINGS

These and other features of the invention will become more apparent from the following description in which reference is made to the appended drawings wherein:

FIG. 1 show various sequences and vectors. FIG. 1A shows the nucleotide of Arabidopsis GLK1 (SEQ ID NO: 1). FIG. 1B shows the amino acid of GLK1 (SEQ ID NO: 2); FIG. 1C shows a schematic diagram of pHS723-70S-AtGLK1-noster, a vector comprising Arabidopsis GLK1; FIG. 1D shows an identity comparison of several known GLK1 amino acid sequences. From top to bottom: Arabidopsis GLK1 polypeptide (accession No. AAK20120; SEQ ID NO: 2), Triticum GLK1 (accession No. ABL10089; SEQ ID NO: 37), Oryza sativa GLK1 (accession No. AAK50393; SEQ ID NO:38), Maize GLK1 (accession No. AAK50392; SEQ ID NO:36), Arabidopsis GLK2 (accession No. AAK20121; SEQ ID NO: 41), Maize GLK2 (SEQ ID NO:39), Oryza sativa GLK2 (accession No. AAK50394; SEQ ID NO:40); FIG. 1E shows the nucleotide sequence of Maize GLK1 (accession No. AF318580.1; SEQ ID NO:33); FIG. 1F shows the nucleotide sequence of Triticum (accession No. EF105406; SEQ ID NO:34); FIG. 1G shows the nucleotide sequence of Oryza sativa (accession No. AF318581.1; SEQ ID NO:35).

FIG. 2 show data obtained from plants over-expressing GLK. FIG. 2A shows Northern blot of GLK over-expressing lines with GLK 1 as probe; FIG. 2B shows RT-PCR of GLK1 over-expressing lines. The left-most lane is a marker lane; WT refers to wild type Arabidopsis plants; and 2-6 and 5-7 refer to GLK1 over-expressing lines; and C) Left panel: Wild Type Arabidopsis; middle panel Arabidopsis over-expressing GLK1 OE line 5-7; right panel Arabidopsis overexpressing GLK1 OE line 2-6. Plants were grown for 24 days under conditions described in Methods.

FIG. 3 shows a comparison of susceptibility to Fusarium graminearum infection between leaves of GLK1 OE lines and wild type by examination of colonization and growth of F. graminearum on leaves. Leaves were inoculated as described in Methods and examined for infection after 96 hours. FIG. 3A shows Infection of leaves of Arabidopsis NahG plants. The appearance of hyphae/mycelia as well as conidiophores are easily discernable. FIG. 3B shows Infection of leaves of wild-type plants. The appearance of hyphae/mycelia as well as conidiophores are easily discernable. FIG. 3C shows Infection of leaves of plants over-expressing GLK1 (line 5-7). Note the lack of infection even after 96 hours.

FIG. 4 shows RT-PCR confirmation of a subset of highly regulated genes identified by Affymetrix analyses. Lanes: WT, wild type; A, GLK1 OE line 2-6; B, GLK1 OE line 5-7

FIG. 5 show the nucleotide and amino acid sequences of At1g77960. FIG. 5A shows the nucleotide sequence (SEQ ID NO: 3) of At1g77960. FIG. 5B shows the amino acid (SEQ ID NO: 4) of At1g77960.

FIG. 6 shows a schematic diagram of the wheat GLK1 vector, pAct-TaGLK1-nos.

FIG. 7 shows Phox-bem1p (PB1) and ASN1 transcripts in TaGLK1oe and AtGLK1oe Arabidopsis lines detected using RT-PCR.

FIG. 8 shows overexpression of TaGLK1 in T1 plants of 3 lines of transgenic wheat, determined using RT-PCR.

FIG. 9 shows TaGLK1 oe vs null labeled.

FIG. 10 shows Fusarium Head Blight (FHB) in TaGLK1oe wheat 14 dpi of F. graminearum. Maturing wheat heads at 50% anthesis were spray individually with 5 ml of F. graminearum inoculum containing 50,000 macroconidia spores/ml. The sprayed plants were placed in a Conviron E15 environmental chamber equipped with a fine water mist that is activated at 30 min intervals and set at 25° C., 16 h light at 200 μEm⁻²

FIG. 11 shows ASN1 expression in Arabidopsis plants overexpressing TaGLK1oe and AtGLK1oe, determined using RT-PCR.

FIG. 12 shows B73 and AtGLK1 OE plants.

FIG. 13 shows vectors for transforming various plants. FIG. 13A shows pTF 101 1 vector for transformation of maize, rice and soybeans with At GLK1. FIG. 13B shows the vector pTF101.1-70S-AtGLK1-nos

DETAILED DESCRIPTION

The present invention relates to expressing GLK in plants. More specifically, the invention provides GLK1 molecules for conferring disease resistance to plants, increasing nitrogen status in plants, or both.
The present invention provides, in part, GLK1 nucleic acid molecules and polypeptides that can be used to confer resistance to a pathogen in a plant, increase nitrogen status in plants, or both confer resistance to a pathogen in a plant and increase nitrogen status in plants.
Also provided are methods of detecting disease resistance genes and plants. The results described herein indicate that overexpression of GLK1 can alter gene expression networks resulting in the upregulation of defense related genes as well as conferring constitutive resistance to colonization by a pathogen in a plant, for example, F. graminearum in Arabidopsis.
The present invention provides a method for a method of conferring resistance to a pathogen in a plant, comprising:
i) providing a plant comprising a nucleic acid molecule encoding a GLK1 polypeptide, the nucleic acid molecule operatively linked to a non-native regulatory region,
ii) growing the plant and expressing the GLK1 polypeptide,
wherein the GLK1 polypeptide is over-expressed from about 2 to about 100 fold relative to GLK1 produced within a wild type plant, and whereby the amount of over-expressed GLK1 is sufficient to confer resistance to the pathogen in the plant.
The present invention also provides a pathogen resistant plant comprising a GLK1 nucleic acid molecule that encodes a GLK1 polypeptide, wherein expression of the GLK1 produces at least 2 fold greater GLK1 polypeptide relative to a wild-type plant.
As described herein, it has been observed that plants that over-express GLK1 exhibit resistance to one or more than one pathogen. Without wishing to be bound by theory, this resistance to one or more than one pathogen by a plant in which GLK1 has been over-expressed, may be a result of the direct over-expression of GLK1, or it may arise indirectly from the over-expression of GLK1. For example, over-expression of GLK1 has been shown to induce an assortment of other nucleotide sequences as shown in FIGS. 4, 7 and 11, and Tables 1-3, and these other nucleotide sequences may result in pathogen resistance.
A “pathogen” is an organism, for example a micro-organism, that causes disease. Plant pathogens include, without limitation, Alternaria spp., Ascochyta spp., Aspergillus spp., Botrytis spp., Cercospora spp., Colletotrichum spp., Diplodia spp., Erwinia spp., Erysiphe spp., Fusarium spp. (e.g., Fusarium graminearum), Gaeumanomyces spp., Helminthosporium spp., Macrophomina spp., Magnaporthe spp., Mycosphaerella spp., Nectria spp., Peronospora spp., Phakopsora spp. (e.g., Phakopsora pachyrhizi or Phakopsora meibomiae), Phoma spp., Phymatotrichum spp., Phytopthora spp. (e.g., Phytopthora infestans), Plasmopara spp., Podosphaera spp., Pseudomonas spp., Puccinia spp. (Puccinia graminis, Puccinia recondite, Piccinia triticinia), Puthium spp., Pyrenophora spp., Pyricularia spp., Pythium spp., Rhizoctonia spp., Scerotium spp., Sclerotinia spp. (e.g., Sclerotinia sclerotium), Septoria, Thielaviopsis spp., Uncinula spp., Venturia spp., Verticillium spp., or Xanthomonas spp., etc. and Phytopthora infestans.
Plants that may be infected by one or more of such pathogens include, without limitation, Arabidopsis, canola, Brassica spp., maize, tobacco, alfalfa, potato, tomato ginseng, pea, oat, rice, soybean, wheat, barley, sunflower, cotton, horticultural plants, or other plant.
The present invention also provides a method of increasing the nitrogen status in a plant, the method comprising:
i) introducing a nucleic acid molecule encoding a GLK1 polypeptide or fragment or variant thereof into a plant cell to produce a transgenic plant cell, such that the nucleic acid molecule is positioned for expression in the transgenic plant cell; and
ii) growing a transgenic plant from the transgenic plant cell,
wherein the GLK1 polypeptide or fragment or variant thereof is over-expressed relative to wild type GLK1 at least 2 fold, whereby the 2 fold over-expression is sufficient to confer resistance to the pathogen in the plant.
The present invention also provides a plant, or a plant cell, that is characterized as having increase nitrogen status. The plant with increased nitrogen status comprising a GLK1 nucleic acid molecule that encodes a GLK1 polypeptide, wherein expression of the GLK1 produces at least 2 fold greater GLK1 polypeptide relative to a wild-type plant. The increase in nitrogen status determined by comparing the plant comprising the GLK1 nucleic acid molecule with a plant that does not express the GLK1 nucleic acid molecule and determining the levels of expression of ASN1, the colour of the leaves, or both determining the levels of expression of ASN1 and the colour of the leaves. Wherein an increase in the expression of ASN1, a darker green leaf, or both determining the levels of expression of ASN1 and the colour of the leaves, when compared to the plant that does not express GLK1, indicates an increase in nitrogen status.
A GLK1 nucleic acid molecule is any nucleic acid molecule that encodes a GLK1 polypeptide as described herein, or a fragment or variant thereof, providing the fragment or variant thereof when expressed in a plant results in resistance to a pathogen when compared to the same plant when the fragment or variant is not expressed. Non-limiting examples of several GLK1 polypeptide sequences may be found in FIG. 1 and are known in the art, for example but not limited to Rossini L. et al. (2001, Plant Cell 13:1231-1244; which is incorporated herein by reference). Accordingly, a GLK1 polypeptide includes an Arabidopsis GLK1 polypeptide (accession No. AAK20120; SEQ ID NO: 2), Triticum GLK (accession No. ABL10089; SEQ ID NO: 37), Maize GLK1 (accession No. AAK50392; SEQ ID NO:36), Oryza saliva (accession No. AAK50393; SEQ ID NO:38), or a fragment or variant thereof, that is capable of conferring resistance to a pathogen in a plant. A GLK1 nucleic acid molecule includes any nucleotide sequence that may encode the amino acid sequences described above, for example, but not limited to SEQ ID NO's: 2, 36, 37 and 38, and include, but are not limited to Arabidopsis GLK1 (NCBI/GenBank accession No. NM 127617.2 or AY028367; SEQ ID NO: 1), Maize GLK1 (accession No. AF318580.1; SEQ ID NO:33), Triticum (accession No. EF105406 or ABL10089; SEQ ID NO:34), Oryza sativa (accession No. AF318581.1; SEQ ID NO:35).
A “pathogen-resistant”, “pathogen resistance”, “disease-resistant”, or “disease resistance” nucleic acid molecule or polypeptide is any nucleic acid molecule or polypeptide, or fragment or variant thereof, that is capable of conferring resistance to a pathogen or a disease in a plant. Pathogen, or disease resistance may be determined by observation of a unit plant surface area, for example as shown in FIGS. 3A-C, where pathogen infestation of a wild-type plant is compared to a plant that is expressing the nucleotide of the present invention that results in pathogen resistance. Pathogen resistance may also be determined quantitatively by measuring the degree of pathogen infestation for a given area, for example, the number or hyphae per unit plant surface area, for example, a leaf surface area, the length of hyphae per unit plant surface area, the number or size of necrotic lesions or other pathogenic factor, per unit plant surface area, and compare the values between a wild-type plant and a plant expressing the nucleic acid sequence of the present invention. A plant that exhibits a reduction of the number of hyphae per unit plant surface area, the length of hyphae per unit plant surface area, the number or size of necrotic lesions or other pathogenic factor, per unit plant surface area, of from about 2% to about 100% or any amount therebetween, is considered pathogen resistant. For example a plant that exhibits a reduction of from about 5% to about 25% or any amount therebetween, 10% to about 75% or any amount therebetween, 25%-75% or any amount therebetween, or a reduction of 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100%, in the amount of pathogen infestation for a given area, for example, the number or hyphae per unit plant surface area (e.g. leaf surface area), the length of hyphae per unit plant surface area, the number or size of necrotic lesions or other pathogenic factor, per unit plant surface area is considered pathogen resistant. One or more than one nucleic acid molecule encoding one or more than one polypeptide that results in pathogen resistance in a plant may include a nucleic acid sequence encoding GLK1 as described herein (for example SEQ ID NO:1, or a fragment or variant thereof), the GLK1 polypeptide (for example SEQ ID NO:2, or a fragment or variant thereof), or a nucleic one or polypeptide, that is induced as a result of the over-expression of GLK1 in a plant, for example, one or more than one of the molecules described in Tables 1, 2, 3 or FIGS. 4, 7 and 11, herein.
A nucleic acid molecule or polypeptide that is capable of “conferring resistance” to a pathogen in a plant is a molecule, that when present or over-expressed in a plant, plant cell, plant tissue, or seed, protects that plant, plant cell, plant tissue, or seed from infection by a pathogen i.e., prevents or reduces the disease response of the plant, plant cell, plant tissue, or seed to that pathogen, when compared to the disease response of a plant, plant cell, plant tissue, or seed in which the nucleic acid molecule or polypeptide is absent, under-expressed, or present at “wild-type” levels. In some embodiments, such a nucleic acid molecule or polypeptide confers “non-host resistance.” Such a nucleic acid molecule or polypeptide may include a GLK1 molecule or may include one or more of the molecules described in Table 1 or FIG. 3 herein.
By “operatively linked” it is meant that the particular sequences interact either directly or indirectly to carry out an intended function, such as mediation or modulation of gene expression. The interaction of operatively linked sequences may, for example, be mediated by proteins that interact with the operatively linked sequences. A coding region of interest may also be introduced within a vector along with other sequences, typically heterologous, to produce a chimeric construct.
The present invention contemplates a nucleotide sequence encoding GLK1 or a biologically active fragment or variant thereof, wherein the fragment or variant encodes a polypeptide that confers pathogen resistance when over-expressed. For example, without wishing to be bound by theory, the over-expression of GLK1 may confer pathogen resistance, or it may result in increased expression of one or more than one of a second protein, for example, but not limited to those proteins identified in FIGS. 4, 7, 11, Tables 1-3, or a both FIGS. 4, 7, 11 and Tablet-3. The increased expression of one or more of a second protein may result in pathogen resistance, increased nitrogen status, or both pathogen resistance and increased nitrogen status.
In a preferred embodiment, the nucleotide sequence that is over-expressed in a plant comprises SEQ ID NO:1. However, the nucleotide sequence may also include a variant that comprises between about 70% to 100% sequence identity, or any amount therebetween with SEQ ID NO:1, for example between about 80% to 100% sequence identity, or any amount therebetween with SEQ ID NO:1, between about 85% to 100% sequence identity, or any amount therebetween with SEQ ID NO:1, between about 90% to 100% sequence identity, or any amount therebetween with SEQ ID NO:1, for example, but not limited to about 70%, 72,%, 75%, 78%, 80%, 82%, 85%, 87%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or about 100% similarity, or any amount therebetween, including the sequence identity that may be determined between the GLK1 sequences provided in FIG. 1D. Provided that the fragment or variant thereof when over-expressed in a plant confers pathogen or disease resistance to the plant. For example, as reported in Rossini et al (2001, Plant Cell 13:1231-1244), ZmGLK1 and OsGLK1 exhibit 72.3% sequence identity. The degree of sequence identity between different GLK1 sequences is typically greater than that between GLK1 and GLK2 of the same or different organism (e.g. identity between ZmGLK1 and ZmGLK2 is 49.3%, OsGLK1 and OsGLK2 is 58.0%). It is preferred that the variant or fragment of GLK1 comprise regions of high sequence identity that are known between GLK1 polypeptides (see FIG. 1D) including a GARP domain, comprising a putative DNA binding domain (Rossini et al., 2001, Plant Cell 13:1231-1244; and US 2007/0022495; both of which are incorporated herein by reference) that folds as an HLH in PHD computer predictions, with the amino acid sequence PELHRR (amino acid 230 in FIG. 1D) starting at first helix is about 14 amino acids long and separated from a second helix by about a 22 amino acid loop, with the second helix starts at NI/VASHLQ (amino acid 266 in FIG. 1D).
To determine whether a nucleic acid exhibits sequence identity with the sequences presented herein, the sequence may be aligned and % identity determined manually, or an oligonucleotide alignment algorithm may be used, for example, but not limited to BLAST (GenBank URL: ncbi.nlm.nih.gov/cgi-bin/BLAST/; using default parameters: Program: blastn; Database: nr; Expect 10; filter: default; Alignment: pairwise; Query genetic Codes: Standard (1)), BLAST2 (EMBL URL: embl-heidelberg.de/Services/index.html using default parameters: Matrix BLOSUM62; Filter: default, echofilter: on, Expect: 10, cutoff: default; Strand: both; Descriptions: 50, Alignments: 50), or FASTA (using default parameters). Other similar algorithms may be employed to determine sequence identity between two or more amino acid sequences as would be know to one of skill in the art.
The present invention also includes nucleotide sequences encoding polypeptides having GLK1 activity, that is encoding a polypeptide that confers pathogen resistance when over-expressed in a plant. The nucleotide sequence may encode GLK1 for example, SEQ ID NO:1, or a sequence that hybridizes to SEQ ID NO:1, or a complement of SEQ ID NO:1, under stringent hybridization conditions (see Maniatis et al., in Molecular Cloning (A Laboratory Manual), Cold Spring Harbor Laboratory (1982) p 387 to 389; Ausubel, et al. (eds), 1989, Current Protocols in Molecular Biology, Vol. 1, Green Publishing Associates, Inc., and John Wiley & Sons, Inc., New York, at p. 2.10.3; both of which are incorporated herein by reference). Without wishing to be limiting in any manner, representative examples of stringent hybridization conditions include hybridization in 4×SSC at 65° C. for 8-16 hours, followed by one, two or three washes in 0.1×SSC at 65° C. for an hour, or hybridization in 5×SSC and 50% formamide at 42° C. for 8 to 16 hours, followed by one, two or three washes in about 0.5×SSC to about 0.2×SSC at 65° C. for one hour. However, hybridization conditions may be modified in accordance with known methods depending on the sequence of interest (see Tijssen, 1993, Laboratory Techniques in Biochemistry and Molecular Biology—Hybridization with Nucleic Acid Probes, Part I, Chapter 2 “Overview of principles of hybridization and the strategy of nucleic acid probe assays”, Elsevier, N.Y., which is herein incorporated by reference). Generally, but not wishing to be limiting, stringent conditions are selected to be about 5° C. lower than the thermal melting point for the specific sequence at a defined ionic strength and pH, as can be determined by one of skill in the art.
The GLK1 and other nucleic acid molecules and polypeptides described herein may be isolated and produced using standard recombinant and other techniques as described herein or known in the art e.g. see Maniatis et al., in Molecular Cloning (A Laboratory Manual), Cold Spring Harbor Laboratory (1982) p 387 to 389; Ausubel, et al. (eds), 1989, Current Protocols in Molecular Biology, Vol. 1, Green Publishing Associates, Inc., and John Wiley & Sons, Inc., New York, at p. 2.10.3; both of which are incorporated herein by reference).
By “DNA regulatory region” it is meant a nucleic acid sequence that has the property of controlling the expression of a DNA sequence that is operably linked with the regulatory region. Such regulatory regions may include promoter or enhancer regions, and other regulatory elements recognized by one of skill in the art. By “promoter” it is meant the nucleotide sequences at the 5′ end of a coding region, or fragment thereof that contain all the signals essential for the initiation of transcription and for the regulation of the rate of transcription. The promoters used to exemplify the present invention are constitutive promoters that are known to those of skill in the art. However, if tissue specific expression of the gene is desired, for example seed, or leaf specific expression, then promoters specific to these tissues may also be employed. Furthermore, as would be known to those of skill in the art, inducible promoters may also be used in order to regulate the expression of the gene following the induction of expression by providing the appropriate stimulus for inducing expression. In the absence of an inducer the nucleic acid sequence will not be transcribed. Typically the protein factor, that binds specifically to an inducible promoter to activate transcription, is present in an inactive form which is then directly or indirectly converted to the active form by the inducer.
The inducer can be a chemical agent such as a protein, metabolite, growth regulator, herbicide or phenolic compound or a physiological stress imposed directly by heat, cold, salt, or toxic elements or indirectly through the action of a pathogen or disease agent such as a virus. A plant cell containing an inducible regulatory element may be exposed to an inducer by externally applying the inducer to the cell or plant such as by spraying, watering, heating or similar methods. Inducible elements may be derived from either plant or non-plant genes (e.g. Gatz, C. and Lenk, I. R. P., 1998, Trends Plant Sci. 3, 352-358; which is incorporated by reference). Examples, of potential inducible promoters include, but not limited to, teracycline-inducible promoter (Gatz, C., 1997, Ann. Rev. Plant Physiol. Plant Mol. Biol. 48, 89-108; which is incorporated by reference), steroid inducible promoter (Aoyama, T. and Chua, N. H., 1997, Plant J. 2, 397-404; which is incorporated by reference), for example but not limited to a dexamethasone-induced promoter, and ethanol-inducible promoter (Salter, M. G., et al, 1998, Plant Journal 16, 127-132; Caddick, M. X., et al, 1998, Nature Biotech. 16, 177-180, which are incorporated by reference) cytokinin inducible IB6 and CKI1 genes (Brandstatter, I. and Kieber, J. J., 1998, Plant Cell 10, 1009-1019; Kakimoto, T., 1996, Science 274, 982-985; which are incorporated by reference) and the auxin inducible element, DR5 (Ulmasov, T., et al., 1997, Plant Cell 9, 1963-1971; which is incorporated by reference). By using an inducible promoter that is operatively linked to the nucleotide sequence of the present invention encoding GLK1, GLK1 may be selectively expressed. This may be desired if the expression of GLK1 leads to reduced plant growth or development, for example if constitutive expression of GLK1 leads to reduced plant growth or delays plant development.
It is also contemplated that the nucleotide sequence of the present invention may also be operatively linked to a wound inducible (Titarenko E., et al., 1997, Plant Physiol. 115:817-826), or pathogenesis related promoter, for example but not limited to a promoter from the gene encoding any of the pathogenesis related proteins PR1-PR15 (van Loon et al., 1999, Phys Mol. Plant. Pathol., 55:85-97; Durrant W. E., Dong. X 2004, Annu. Rev. Phytopatho; 1. 42:185-209, which are incorporated herein by reference), including PRI (e.g. Payne G., et al., 1988, Plant Mol. Biol. 11:89-94; Payne G et al., 1989 Plant Mol. Biol. 12:595-596), or a promoter from PRIa-PRIg (van Loon et al., 1999, Phys Mol. Plant. Pathol., 55:85-97), PR2 (Ward E. R. 1991, Plant Physiol. 96:390-397), PR3 (Payne G., et al. 1990, PNAS 81:98-102), PR4 (Ward E. R., 1991, Plant Cell, 3:1085-1094), PR5 (Payne G., et al., 1998, Plant Mol. Biol. 11:232-234). Additionally, the nucleotide sequence of the present invention may be operatively linked to a promoter encoding a stress-induced protein, resulting from either an abiotic or biotic stress, for example but not limited to a promoter obtained from a jasmonic acid induced gene, for example NPR1 (Kachroo N. A., et al., 2003, Mol. Plant. Microbe Interact. 16:588-599). By using a stress, or would-inducible promoter that is operatively linked to the nucleotide sequence of the present invention encoding GLK1, GLK1 may be induced under conditions that result in its selective expression. This may be desired if the expression of GLK1 leads to reduced plant growth or development.
A constitutive regulatory element directs the expression of a gene throughout the various parts of a plant and continuously throughout plant development. Examples of known constitutive regulatory elements include promoters associated with the CaMV 35S transcript (Odell et al., 1985, Nature, 313: 810-812), the rice actin 1 (Zhang et al, 1991, Plant Cell, 3: 1155-1165) and triosephosphate isomerase 1 (Xu et al, 1994, Plant Physiol. 106: 459-467) genes, the maize ubiquitin 1 gene (Cornejo et al, 1993, Plant Mol. Biol. 29: 637-646), the Arabidopsis ubiquitin 1 and 6 genes (Holtorf et al, 1995, Plant Mol. Biol. 29: 637-646), tobacco t-CUP promoter (WO/99/67389; U.S. Pat. No. 5,824,872), the HPL promoter (WO 02/50291), and the tobacco translational initiation factor 4A gene (Mandel et al, 1995 Plant Mol. Biol. 29: 995-1004). The term “constitutive” as used herein does not necessarily indicate that a gene under control of the constitutive regulatory element is expressed at the same level in all cell types, but that the gene is expressed in a wide range of cell types even though variation in abundance is often observed.
The chimeric gene construct of the present invention can further comprise a 3′ untranslated region. A 3′ untranslated region refers to that portion of a gene comprising a DNA segment that contains a polyadenylation signal and any other regulatory signals capable of effecting mRNA processing or gene expression. The polyadenylation signal is usually characterized by effecting the addition of polyadenylic acid tracks to the 3′ end of the mRNA precursor. Polyadenylation signals are commonly recognized by the presence of homology to the canonical form 5′ AATAAA-3′ although variations are not uncommon. Examples of suitable 3′ regions are the 3′ transcribed non-translated regions containing a polyadenylation signal of Agrobacterium tumor inducing (Ti) plasmid genes, such as the nopaline synthase (Nos gene) and plant genes such as the soybean storage protein genes and the small subunit of the ribulose-1,5-bisphosphate carboxylase (ssRUBISCO) gene. The 3′ untranslated region from the structural gene of the present construct can therefore be used to construct chimeric genes for expression in plants.
The chimeric gene construct of the present invention can also include further enhancers, either translation or transcription enhancers, as may be required. These enhancer regions are well known to persons skilled in the art, and can include the ATG initiation codon and adjacent sequences. The initiation codon must be in phase with the reading frame of the coding sequence to ensure translation of the entire sequence. The translation control signals and initiation codons can be from a variety of origins, both natural and synthetic. Translational initiation regions may be provided from the source of the transcriptional initiation region, or from the structural gene. The sequence can also be derived from the regulatory element selected to express the gene, and can be specifically modified so as to increase translation of the mRNA.
To aid in identification of transformed plant cells, the constructs of this invention may be further manipulated to include plant selectable markers. Useful selectable markers include enzymes which provide for resistance to an antibiotic such as gentamycin, hygromycin, kanamycin, and the like. Similarly, enzymes providing for production of a compound identifiable by colour change such as GUS (beta-glucuronidase), or luminescence, such as luciferase are useful.
Also considered part of this invention are transgenic plants containing the chimeric gene construct comprising a nucleotide sequence encoding GLK1 as described herein. However, it is to be understood that the nucleotide sequence of the present invention encoding GLK1, may also be combined with a range of regulatory elements for expression within a range of host organisms. Such organisms include, but are not limited to, corn, wheat, barley, oat, tobacco, Brassica, soybean, pea, alfalfa, potato, ginseng, Arabidopsis, horticultural plants, and other plants.
Methods of regenerating whole plants from plant cells are also known in the art. In general, transformed plant cells are cultured in an appropriate medium, which may contain selective agents such as antibiotics, where selectable markers are used to facilitate identification of transformed plant cells. Once callus forms, shoot formation can be encouraged by employing the appropriate plant hormones in accordance with known methods and the shoots transferred to rooting medium for regeneration of plants. The plants may then be used to establish repetitive generations, either from seeds or using vegetative propagation techniques.
The constructs of the present invention can be introduced into plant cells using Ti plasmids, Ri plasmids, plant virus vectors, direct DNA transformation, micro-injection, electroporation, etc. For reviews of such techniques see for example Weissbach and Weissbach, Methods for Plant Molecular Biology, Academy Press, New York VIII, pp. 421-463 (1988); Geierson and Corey, Plant Molecular Biology, 2d Ed. (1988); and Miki and Iyer, Fundamentals of Gene Transfer in Plants. In Plant Metabolism, 2d Ed. DT. Dennis, D H Turpin, D D Lefebrve, D B Layzell (eds), Addison Wesly, Langmans Ltd. London, pp. 561-579 (1997). For Arabidospsis see Clough and Bent (1998, Plant J. 16, 735-743). The present invention further includes a suitable vector comprising the chimeric gene construct.
As described herein, a network of gene products showed increased transcript accumulation as a result of over-expression of GLK1. The most highly accumulated transcripts of this GLK1 ‘regulon’ encodes genes that are associated with plant defense against mainly biotic stress (Table 1, FIG. 4). Without wishing to be bound by theory, the reprogrammed network may have resulted in the development of a non-host resistant phenotype to a pathogen, for example Fusarium graminearum. This therefore identifies a role for GLK1 other than of chloroplast differentiation as is known in the prior art.
The suite of defense related genes that appear to be associated with the GLK1 regulon include genes that are known to respond to reactive oxygen species (ROS) as well as genes that function in detoxification such as cytochrome p450 (CYP81 family) and MATE efflux proteins. MATE efflux proteins belong to a family of multidrug and toxin extrusion efflux transporters in plants and microbes (Brown, M. H. et al. 1998, Mol. Microbiol. 31:393-395). One member, ALF5, has been shown to be required for resistance to toxins in Arabidopsis (Diener, A. C., et al., 2001, Plant Cell 13:1625-1637). In addition, the eds5 mutation in Arabidopsis encodes a MATE efflux protein that is strongly induced by pathogens, SA application and UV-light exposure (Nawrath, C., et al., 2002, Plant Cell 14: 275-286). The upregulation of MATE efflux protein is consistent with the F. graminearum resistant phenotype observed here as the trichothecene toxin deoxnivalenol (DON) which is produced by the pathogen has been shown to be a factor in infection (Harris, L. J., et al., 2001, Physiol. Mol. P1. Path. 58:173-181).
A pathogen related gene PR10 was also identified to be highly upregulated. PR10 has been observed to be upregulated by infection with pathogens (Riggleman, R. C., 1985, Plant Mol. Biol. 4:81-86) and has been observed to possess antimicrobial activity (C-J. Park, et al., 2004, Plant J. 37:186-198). One of the most highly accumulating transcripts in the GLK1 regulon however, encodes an unknown protein containing a Phox/Bem 1 or PBI domain. The function of this PBI domain containing protein is not known. However, PBI domains are involved in protein-protein interactions and are involved in the activation of NADPH oxidase in responses to ROS as well as suppressing cell death (Simon-Plas, F., et al., 2002, Plant J., 31:137-147; Torres, M. A., et al., 2005, Nature Genetics 37:1130-1134), in MEKK kinase signaling and have also been observed to be upregulated in response to pathogen (Narusaka, Y., et al., 2004, Mol. Plant. Microbe Inter. 17:749-762).
Included in the GLK1 regulon are cinnamyl alcohol dehydrogenase (CAD), and ELI3-2, an aromatic alcohol dehydrogenase. CAD has bee demonstrated to be upregulated in response to pathogen and is implicated in defense lignin biosynthesis, as well as non-host defense in wheat. ELI3-2 encodes an aromatic alcohol dehydrogenase that is upregulated in association with local infection sites. Its expression is has been shown to be dependent on RPM1 expression. GLK1 over-expression also upregulated isochorismate synthase (At1g18870) transcription. Another highly accumulating gene transcript in GLK1 OE is At2g41050, a PQ-loop repeat protein homologous to cystinosin (a lysosomal cystine transporter).
As shown herein, it is possible that GLK1 over expression (OE) may directly transactivate a number of defense related genes. The phenotype of GLK1 OE may be the result of up-regulation, down-regulation, or both up-regulation and down-regulation, of these genes to prepare (prime) the plant for pathogen invasion. Priming is a phenomenon that preconditions the plant to stress. Therefore, one or more than one of the nucleic acid sequences identified in FIG. 3 that exhibit up-regulation, for example of about 2 fold or more, in expression, down-regulation, for example, of 2 fold or more in expression, or both up-regulation and down-regulation, for example, of 2 fold or more, may be used to screen a plant of interest to determine if the plant exhibits pathogen resistance. Example of sequences that may be up or down regulated include those listed in Tables 1-3.
The present invention therefore provides a genetic construct comprising a regulatory region operably linked to a nucleic acid molecule encoding a GLK1 polypeptide or fragment or variant thereof. The nucleic acid molecule may comprise a nucleic acid sequence as set forth in SEQ ID NO: 1, a nucleotide sequence that exhibits from about 70% to about 100% sequence identity with the sequence of SEQ ID NO:1, or a nucleotide sequence that hybridizes to the nucleic acid sequence set forth in SEQ ID NO: 1 under conditions of high stringency. The GLK1 polypeptide may comprise an amino acid sequence set forth in SEQ ID NO: 2. The present invention also pertains to a host cell, a transgenic plant, or a transgenic plant seed, comprising the genetic construct as just described, the transgenic plant is characterized as being resistant to a pathogen, as exhibiting increased nitrogen status, or both resistant to a pathogen and exhibiting increased nitrogen status.
The present invention also provides a method of detecting a candidate pathogen-resistance gene in a plant, comprising, comparing the gene expression profile of a transgenic plant over-expressing a nucleic acid molecule encoding a GLK1 polypeptide or fragment or variant thereof, with the gene expression profile of a wild-type plant; and determining the genes over-expressed in the transgenic plant compared to the wild-type plant, wherein one or more than one of the genes over-expressed in the transgenic plant are candidate pathogen-resistance genes. The present invention also provides a method as just described wherein the GLK1 polypeptide is selected from the group consisting of an Arabidopsis GLK1, a wheat GLK1 polypeptide, a maize GLK1 polypeptide, a rice GLK1 polypeptide, and a soy bean GLK1 polypeptide.
The present invention also pertains to a method of detecting a plant exhibiting resistance to a pathogen, the method comprising:
i) determining the expression levels of a GLK1 nucleic acid molecule or polypeptide or fragment or variant thereof in a test plant;
ii) determining the expression levels of the GLK1 nucleic acid molecule or polypeptide or fragment or variant thereof in a control plant; and
iii) comparing the expression levels in the test plant to the expression levels in the control plant,
wherein the over-expression of the GLK1 nucleic acid molecule or polypeptide or fragment or variant thereof in the test plant compared to the control plant is indicative of the test plant being a plant exhibiting resistance to a pathogen. The test plant may be selected from the group consisting of Arabidopsis, canola, Brassica spp., maize, tobacco, alfalfa, potato, ginseng, pea, oat, rice, soybean, wheat, barley, sunflower, and cotton.
Furthermore the present invention includes a method of detecting a plant exhibiting resistance to a pathogen, the method comprising:
i) determining the expression levels of one or more than one of the nucleic acid molecule or polypeptide set out in Table 1 or in FIG. 3, or a fragment or variant thereof, in a test plant;
ii) determining the expression levels of the one or more of the nucleic acid molecule or polypeptide set out in Table 1 or in FIG. 3, or a fragment or variant thereof, in a control plant; and
iii) comparing the expression levels in the test plant to the expression levels in the control plant,
wherein the over-expression of the one or more of the nucleic acid molecule or polypeptide, reduced expression of the one or more of the nucleic acid molecule or polypeptide, or both over-expression of the one or more of the nucleic acid molecule or polypeptide and reduced expression of the one or more of the nucleic acid molecule or polypeptide, set out in Table 1 or in FIG. 3, or a fragment or variant thereof, in the test plant compared to the control plant is indicative of the test plant being a plant exhibiting resistance to a pathogen. The test plant may be selected from the group consisting of Arabidopsis, canola, Brassica spp., maize, tobacco, alfalfa, potato, ginseng, pea, oat, rice, soybean, wheat, barley, sunflower, and cotton.
The method of detecting a plant (a first plant) that exhibits resistance to a pathogen as described above may comprise detecting a nucleic acid molecule that is expressed at least about 2 fold higher when compared to the expression of the same nucleic acid molecule within a wild-type plant (a second plant). Preferably, the nucleic acid molecule is expressed from about 5 to about 150 fold higher, or any amount therebetween, when compared to the expression of the same nucleic acid molecule within a wild-type plant (a second plant). For example, the nucleic acid molecule may be expressed from about 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, 16, 18, 20, 22, 25, 28, 30, 35, 40, 45, 50, 55, 60, 665, 70, 75, 80, 90, 100, 110, 120, 130, 140, 150 fold higher, or any amount therebetween, when compared to the expression of the same nucleic acid molecule within a wild-type plant (a second plant). This difference in expression may be determined using any method that can differentiate between the levels of expression between the first and second plant, for example but not limited to northern analysis, dot blot analysis, quantitative PCR, a microarray, or other method as would be known to one of skill in the art. Examples of nucleic acids that may be used within this method include those that encode, but are not limited to one or more than one of the following: a putative Mn superoxide dismutase (At3g56350), a glycosyl hydrolase family 1 protein (At3g60140), a Phox/Bem1p domain protein (At1g77960), proline-rich extensin-like family protein (At5g49080), a Bet v I allergen family protein (At1g35310), a trypsin, protease inhibitor family protein (At1g73260), golden2-like transcription factor (GLK1; At2g20570), cinnamyl-alcohol dehydrogenase family protein (At1g09500), a hydroxyproline-rich glycoprotein family protein (At1g77970), a glutamine-dependent asparagine synthetase 1 (At3g47340), a major latex-related protein (At4g23680), a putative mannitol dehydrogenase (ELI3-2; At4g37990), a glycosyl hydrolase family 1 protein (At3g09260), a lactoylglutathione lyase family protein/glyoxalase I family protein (At1g80160), Lactoylglutathione lyase (At1g80160), Isochorismate synthase (At1g18870), a cystinosin PQ-loop repeat protein (At2g41050), MATE efflux protein (At4g21910), Galactinol synthase (At1g09350), or a combination of any of these proteins. Additional nucleic acid markers may also be used as listed in Table 1. It is also contemplated that the proteins encoded by these markers may be used to determine expression of the encoded proteins listed in Table 1. The levels of protein expression may be determined using standard methods including PAGE, staining and scanning the gel, or antibody based techniques, for example ELISA, or diffusion assays as would be known within the art.
Also provided is a method of detecting a plant exhibiting resistance to a pathogen, the method comprising:
i) determining the expression levels of a At1g77960, At2g41050, or both At1g77960 and At2g41050 nucleic acid molecule or polypeptide or fragment or variant thereof in a test plant;
ii) determining the expression levels of the At1g77960, At2g41050, or both At1g77960 and At2g41050 nucleic acid molecule or polypeptide or fragment or variant thereof in a control plant; and
iii) comparing the expression levels in the test plant to the expression levels in the control plant,
wherein the over-expression of the At1g77960, At2g41050, or both At1g77960 and At2g41050 nucleic acid molecule or polypeptide or fragment or variant thereof in the test plant compared to the control plant is indicative of the test plant being a plant exhibiting resistance to a pathogen. The test plant may be selected from the group consisting of Arabidopsis, canola, Brassica spp., maize, tobacco, alfalfa, potato, ginseng, pea, oat, rice, soybean, wheat, barley, sunflower, and cotton.
Affymetrix microarray analyses indicated that over-expression of wheat GLK1 (TaGLK1 Accession No. ABL10089) in Arabidopsis shows the regulation of similar genes as in AtGLK1 OE in Arabidopsis (Tables 1-3).
As shown in FIG. 7, Phox-bem1p (PB1) and ASN1 transcripts are expressed in TaGLK1oe and AtGLK1oe Arabidopsis lines. This data validates the microarray data presented in Table 3 with the genes At1g77960 (PB1) and At3g47340 (ASN1). Taken together, these analyses indicate that TaGLK1 OE is capable of reprogramming identical networks of gene expression as that observed using AtGLK1 OE.
Transgenic wheat (Triticum aestivum cv Fielder) over-expressing wheat TaGLK1 (FIG. 8) shows enhanced resistance to the spread of lesions in wheat leaves inoculated with F. graminearum (FIG. 9) and in the development of Fusarium Head Blight (FHB) symptoms in developing wheat heads (FIG. 10).
As described herein, overexpression of AtGLK1 and TaGLK1 in Arabidopsis also resulted in the upregulation of the glutamine dependent asparagine synthetase 1 ASN1 transcripts (Table 1-3; FIG. 11). Increases in endogenous levels of ASN1 have been suggested to be associated with higher nitrogen status (Tian et al, 2007, J Biol Chem 282: 18532-18541). Over-expression of bacterial ASN1 is known to enhance the nitrogen status in Arabidopsis and soybean sink tissues (Lam et al, 2003, Plant Physiol 132: 926-935; Wan et al, 2006, Plant Biol (Stuttgart) 8: 271-276), enhanced vegetative growth (Giannino et al, 2007, Euphytica DOI 10.1007/s10681-007-9506-3) and increased synthesis of inulin (Sobolev et al, J Agric Food Chem 55, 10827-10831). The over-expression of AtGLK1 in maize cv. Hii2 produced transgenic maize that had darker green leaves when grown under midwinter (early December, Latitude 46N) greenhouse conditions suggestive of enhanced nitrogen utilization (FIG. 12). This demonstrating that over-expression of At and TaGLK1(s) presents an opportunity to modify nitrogen status in crop plants.
Therefore the present invention also provides a method of increasing the nitrogen status in a plant, the method comprising:
i) introducing a nucleic acid molecule encoding a GLK1 polypeptide or fragment or variant thereof into a plant cell to produce a transgenic plant cell, such that the nucleic acid molecule is positioned for expression in the transgenic plant cell; and
ii) growing a transgenic plant from the transgenic plant cell,
wherein the GLK1 polypeptide or fragment or variant thereof is over-expressed relative to wild type GLK1 at least 2 fold, whereby the 2 fold over-expression is sufficient to confer resistance to the pathogen in the plant.
The present invention also provides a plant, or a plant cell, that is characterized as having increase nitrogen status. The plant with increased nitrogen status comprising a GLK1 nucleic acid molecule that encodes a GLK1 polypeptide, wherein expression of the GLK1 produces at least 2 fold greater GLK1 polypeptide relative to a wild-type plant. The increase in nitrogen status may be observed by comparing the plant comprising the GLK1 nucleic acid molecule with a plant that does not express the GLK1 nucleic acid molecule and determining the levels of expression of ASN1, the colour of the leaves, or both. Wherein an increase in the expression of ASN1, a darker green leaf, or both, when compared to the plant that does not express GLK1, indicates an increased nitrogen status.
Furthermore, the present invention provides a macroarray, or a microarray comprising, but are not limited to, one or more than one nucleic acid that encode: a putative Mn superoxide dismutase (At3g56350), a glycosyl hydrolase family 1 protein (At3g60140), a Phox/Bem1p domain protein (At1g77960), proline-rich extensin-like family protein (At5g49080), a Bet v I allergen family protein (At1g35310), a trypsin, protease inhibitor family protein (At1g73260), golden2-like transcription factor (GLK1; At2g20570), cinnamyl-alcohol dehydrogenase family protein (At1g09500), a hydroxyproline-rich glycoprotein family protein (At1g77970), a glutamine-dependent asparagine synthetase 1 (At3g47340), a major latex-related protein (At4g23680), a putative mannitol dehydrogenase (ELI3-2; At4g37990), a glycosyl hydrolase family 1 protein (At3g09260), a lactoylglutathione lyase family protein/glyoxalase I family protein (At1g80160), Lactoylglutathione lyase (At1g80160), Isochorismate synthase (At1g18870), a cystinosin PQ-loop repeat protein (At2g41050), MATE efflux protein (At4g21910), Galactinol synthase (At1g09350), or a combination thereof. The microarray may be used to determine if a plant of interest exhibits pathogen resistance.
The present invention also provides a macroarray, or a microarray comprising one or more than one of the nucleic acid molecule or polypeptide set out in Table 1 or in FIG. 3, or a fragment or variant thereof. The microarray may be used to determine if a plant of interest exhibits pathogen resistance.
For macroarrays, the following non-limiting method may be used where nucleic acid fragments (with known sequence information) are spotted onto a membrane, for example a nylon membrane (Hybond N, Amersham Pharmacia) in a way to ensure that the location of each DNA fragment is known. The arrayed DNA are fixed to the membrane and hybridized to a labelled probe as described by Sambrook et al., 1989, Molecular Cloning, 2^nded., Cold Spring Harbor Laboratory Press. Total RNA is isolated from a test plant (for example, according to the method of Chang et al., Plant Mol. Biol. Rep., 1993, 11:113-116), and Poly A+ mRNA isolated. Labelled cDNA is produced by reverse transcription (for example using SuperScript II Rnase H⁻ reverse transcriptase (GIBCO BRL). The labelled cDNA probes are hybridized to the DNA present in the array and detected using any suitable method, for example, fluorescence, or autoradiography.
Microarrays may be produced using methods known in the art (for example, as described at the following URL: affymetrix [dot] com/technology/tech_probe [dot] html). DNA fragments may also be directly spotted onto glass slides. Labelled probes, for example, fluorescently labelled cDNA probes can be synthesized according to the methods outlined at the following URL: uhnres [dot] utoronto [dot] ca/services/microarray/protocols/Pro_RT [dot] html, using poly A⁺ RNA isolated from the appropriate plant tissue, as described above (in DNA macroarrays), and hybridized to the microarray using standard procedures: uhnres [dot] utoronto [dot] ca/services/microarray/protocols/Pro_hybridization [dot] html. Differentially labeled cDNA probes may be used if desired (eg. cDNA produced from two different sources of RNA labeled with Cyanine 3 and Cyanine 5, respectively). The hybridized slides or arrays are then scanned.
The present invention discloses the following sequences:


SEQ ID NO:	Name

1	GLK1 Arabidopsis
	nucleic acid
2	GLK1 Arabidopsis
	amino acid
3	At1g77960 nucleic acid
4	At1g77960 amino acid
5	GLK1-5′Xba primer
6	GLK1-3′Kpn primer
7	At2g20570 5′ primer
8	At2g20570 3′ primer
9	At1g09350 5′ primer
10	At1g09350 3′ primer
11	At1g09500 5′ primer
12	At1g09500 3′ primer
13	At1g35310 5′ primer
14	At1g35310 3′ primer
15	At1g73260 5′ primer
16	At1g73260 3′ primer
17	At1g77960 5′ primer
18	At1g77960 3′ primer
19	At1g80160 5′ primer
20	At1g80160 3′ primer
21	At3g0250 5′ primer
22	At3g0250 3′ primer
23	At3g60140 5′ primer
24	At3g60140 3′ primer
25	At4g37990 5′ primer
26	At4g37990 3′ primer
27	At1g18870 5′ primer
28	At1g18870 3′ primer
29	At2g14610 5′ primer
30	At2g14610 3′ primer
31	5′ primer
32	3′ primer
33	GLK1 Maize nucleic acid
34	GLK1 Wheat nucleic acid
35	GLK1 Rice nucleic acid
36	GLK1 Maize amino acid
37	GLK1 Wheat amino acid
38	GLK1 Rice amino acid
39	GLK2 Maize amino acid
40	GLK2 Rice amino acid
41	AtGLK2 amino acid
42	At1g02450 5′ primer
43	At1g02450 3′ primer
44	primer to TaGLK1 5′
45	primer to TaGLK1 3′
46	At2g415050 5′ primer
47	At2g415050 3′ primer
48	At3g28740 5′ primer
49	At3g28740 3′ primer
50	At1g18870 5′ primer
51	At1g18870 3′ primer
52	At4g21910 5′ primer
53	At4g21910 3′ primer
54	At3g47340 (ASN1) 5′
	primer
55	At3g47340 (ASN1) 3′
	primer
56	ABC10089 (TaGLK1) 5′
	primer
57	ABC10089 (TaGLK1) 3′
	primer

The present invention will be further illustrated in the following examples.

EXAMPLES

Example 1

Materials and Methods

Growth of plant material: Arabidopsis thaliana (ecotype Columbia) seeds were germinated in soil in 5 cm pots. Seedlings were grown in Conviron E15 chambers at 22° C., 16 hour days at a light intensity of 150 μEm⁻². Fertilization was carried once weekly with 20:20:20: NPK.
RNA isolation: Leaf (2.5 cm rosette) tissues were collected 4 hours after the onset of light period. One gram quantities of leaf (rosette stage, 3 cm long) tissues were collected and frozen in liquid N₂. Total RNA was isolated using Trizol™, P/N 15596-018 (Invitrogen, Burlington, ON, Canada) according to manufacturer's instructions. RNA quality was assessed by electrophoresis on formaldehyde gels.
Cloning of AtGLK1 (AT2G20570): RT-PCR was performed, using total RNA and Superscript III (Invitrogen, Burlington, ON, Canada). First strand synthesis reactions were carried for 1 ug of the isolated RNA with oligo dT (T₂₀VN) and Superscript III (Invitrogen 18080-044). The following primers:
(SEQ ID NO: 5)

GLK1 5′ Xba1-TATATTTGGATCTAGAAGTGAAG;

(SEQ ID NO: 6)

GLK1 3′ Kpn1-TTGGTTCTCGGGTACCTACGG

were used to amplify the resultant cDNA fragment. The PCR fragment was subcloned into pGEM-t-easy (Promega, Madison Wis.) and transformed into E. coli DH5α and the insert was confirmed by sequencing using m13 forward and reverse primers. The fragment was excised with Xba and Kpn1 (introduced by the GLK1 primers) and ligated into pHS723 already containing the 70S-nos-terminator cassette to create pHS723-GLK1. pHS723 contains a NPTII-GUS fusion to allow confirmation of selected plants with GUS (Nair, R. B., et al., 2000, Plant Physiol. 123:1623-1634).
Overexpression of Arabidopsis with GLK1: pHS723-GLK1 was introduced into Agrobacterium tumafaciens (strain GV3109 pmp90) and the resulting transformant was used to infect Arabidopsis by the floral dip method (Clough, S. J., Bent, A. F., 1998, Plant J. 16:735-743). Seeds from T_oplants were collected, sterilized and plated on MS media containing 50 μg/ml kanamycin and transformed plants selected as described (Clough, S. J., Bent, A. F., 1998, Plant J. 16:735-743). Plantlets were transferred to soil and GUS staining was used to further confirm integration of the plasmid. GUS stain was also used as a marker to obtain homozygous T₂plants with single insertions. All further experiments were carried out with T₃plants.
Arabidopsis Gene Chip Analyses: Affymetrix Gene Chip hybridization and analyses were carried as per manufacturer's protocols at the McGill University and Genome Quebec Innovation Centre (Montreal), using the Affymetrix Arabidopsis ATH1-121501 Gene Chip representing 24,000 genes. For each replicate, 10 ug of total RNA adjusted to a concentration of 1 μg/ul in DMPC water was used. The data was MAS5.1 mean scale normalized. Microarray analyses were carried out as describe in Savitch et al, (2007, Biochem Biophys Res Comm 359: 234-238). Affymetrix Microarray data were analysed by normalization and T-tests using FlexArray program (see URL: genomequebec [dot] mcgill [dot] ca/FlexArray/).
Validation of the microarray data for selected genes was carried out by RT-PCR as described above with the following primers:

	(SEQ ID NO: 7)
	At2g20570: 5′-TCGGGGAAAGGTGAAGAAGTCGT,
	(SEQ ID NO: 8)
	3′-CAGCCATGGCCTCGTCAATACA;

	(SEQ ID NO: 9)
	At1g09350: 5′-ATGGCACCTGAGATGAACAACAAG,
	(SEQ ID NO: 10)
	3′-GCAAACATATAAGGTACAAAGAGG;

	(SEQ ID NO: 11)
	At1g09500: 5′-CAACCACATGCACACTAATTC,
	(SEQ ID NO: 12)
	3′-CCATTGGCTGAAGGAGTCTCG;

	(SEQ ID NO: 13)
	At1g35310: 5′-CAACATGCCTCCAAAGCCACTC,
	(SEQ ID NO: 14)
	3′-ACGCATACAATAACTCTCCCACAC;

	(SEQ ID NO: 15)
	At1g73260: 5′-CTATCAAGCCGCCTCACCTA,
	(SEQ ID NO: 16)
	3′-CTCACCGACCCGCCAGTA;

	(SEQ ID NO: 17)
	At1g77960: 5′-ATCGCCGTTGTTGAGTCCTTTATG,
	(SEQ ID NO: 18)
	3′-AGTATTTGTGCCCCTTTGTTCAGC;

	(SEQ ID NO: 19)
	At1g80160: 5′-GATGGCGCGTGGTTGTTT,
	(SEQ ID NO: 20)
	3′-GGGGAGGCTATCGCAGTTG;

	(SEQ ID NO: 21)
	At3g02500: 5′-CATCTCTAGTACTGCCTCTTCTCC,
	(SEQ ID NO: 22)
	3′-TGTGTAACCAATATCCCAAAATG;

	(SEQ ID NO: 23)
	At3g60140: 5′-TGCCTCGGCGTTTCAGTA,
	(SEQ ID NO: 24)
	3′-TCTCCGACGACCGATTTCT;

	(SEQ ID NO: 25)
	At4g37990: 5′-CGCCGCACCGCTCCTCTG,
	(SEQ ID NO: 26)
	3′-GCGGTACCTAACGTCGGCTTTCTC;

	(SEQ ID NO: 27)
	At1g18870: 5′-AGTGCCCCAGGTCGAGTTTGATG,
	(SEQ ID NO: 28)
	3′-TCCCGCCTTTCTTTAGTTGATTGG;

	(SEQ ID NO: 29)
	At2g14610: 5′-CTCAAGATAGCCCACAAGA,
	(SEQ ID NO: 30)
	3′-AAGGCCACATATTTTACAT

	(SEQ ID NO: 42)
	At1g02450: 5′-TACAATTATTCCCTAGAGACCAT,
	(SEQ ID NO: 43)
	3′-TACAATTATTCCCTAGAGACCAT

	(SEQ ID NO: 46)
	At2g41050 5′-TATAAACCCGTCGACTCATTCATT
	(SEQ ID NO: 47)
	3′-GGGCAAGTCATTCATATCTAAGGA

	(SEQ ID NO: 48)
	At3g28740 5′-CGGCCGCGTAAACTAAACCTA
	(SEQ ID NO: 49)
	3′-TCCCGGCAAGTATCATAACAAGTA

	(SEQ ID NO: 50)
	At1g18870 5′-AGTGCCCCAGGTCGAGTTTGATG
	(SEQ ID NO: 51)
	3′-TCCCGCCTTTCTTTAGTTGATTGG

	(SEQ ID NO: 52)
	At4g21910 5′-GTTGGCACTACCGGCGATACTTA
	(SEQ ID NO: 53)
	3′-ACACCCAACCGGAATACCAACAAC

	(SEQ ID NO: 54)
	At3g47340(ASN1) 5′-GTGGAATACTTGCCGTGTTAGGAT
	(SEQ ID NO: 55)
	3′-CACGAGCCACCATGAAAGAGTTAT

	(SEQ ID NO: 56)
	ABL10089(TaGLK1) 5′-CCGCGGGCGTCGAGGAGTTG
	(SEQ ID NO: 57)
	3′-CGGCGGCATACATCTGTCGTCGTT

Fusarium graminearum Infection Bioassay: Assessment of Susceptibility to Fusarium graminearum:

Fungal challenge of Arabidopsis leaves with F. graminearum strain DOAM233423 was carried out essentially as described by Makandar et al (Makandar, R., et al., 2006, Mol. Plant. Microbe Interact. 19:23-129) with the following modifications. Two centimeter rosette leaves from four to 5 weeks old Arabidopsis seedlings grown as described above were used for fungal inoculation. A fungal inoculum containing fifty-thousand spores per ml was infiltrated into the abaxial surface of leaves with a needle-less syringe. The pathogen-inoculated plants were covered with a clear plastic dome to maintain high humidity. The disease spread was recorded 48 hrs and 96 hrs post inoculation by Trypan-blue (TB) stain. After destaining, the leaves were stored in 60% glycerol and examined for presence of mycelia under the light microscope.

Example 2

Overexpression of GLK1

To study the effect of GLK in gene regulation, GLK1 and GLK2 were studied in Arabidopsis. More specifically, Arabidopsis GLK 1 cDNA (SEQ ID NO: 1; FIG. 1A), regulated by the tandem 35S promoter (FIG. 1C), was inserted into the transformation vector pHS723 (Nair et al., 2000) to produce the plasmid pHSGLK1. The plasmid was introduced into Arabidopsis by the floral dip method (Clough and Bent, 1998, Plant J. 16:735-743) and two homozygous independent transgenic lines (2-6 and 5-7) exhibiting increased accumulation of AtGLK1 transcripts i.e., over-expressing AtGLK1 were obtained, as confirmed by Northern blotting using full length AtGLK1 (SEQ ID NO:1) and RT-PCR (FIG. 2A-B) using the following primers:

	(SEQ ID NO: 31)
	5′ primer-tcggggaaaggtgaagaagtcgt

	(SEQ ID NO: 32)
	3′ primer-cagccatggcctcgtcaataca

Seedlings (T5) of both GLK1 OE lines were approximately 5 days slower in growth than the wild type under conditions specified in the Methods section (FIG. 2C).

Example 3

Analysis of Disease Phenotypes

The ability of the two independent GLK1 over-expressing lines to respond to pathogenesis was studied using the hypersensitive response test and by resistance to Fusarium graminearum infection.
The hypersensitive response was induced by challenging the leaves with Pseudomonas syringae. Both GLK1 over-expressing lines were able to initiate a hypersensitive response much more rapidly than wild type.
The leaves of transgenic Arabidopsis over-expressing GLK1 were also challenged with a virulent strain of F. graminearum conidia (Chen et al, 2006) and compared to both the wild type (WT) and an Arabidopsis line over-expressing the NahG transgene. The ability of the leaves of the GLK1 over-expressing line to inhibit mycelial growth of Fusarium graminearum was dramatic and is illustrated in FIGS. 3A-3C, which show comparisons between NahG (a SA impaired line that has lost its ability to develop SAR and is susceptible to infection (Gaffney, T., et al. 1993, Science 261:754-756), the wild type (WT) and the GLK1 OE (5-7) lines. The responses to infection were followed over a time course. At four days post inoculation, the presence of the highest density of hyphae and conidiophores was observed on the NahG line (FIG. 3A) followed by the WT line (FIG. 3B). No mycelial growth of the fungus was observed on the leaves of the GLK1 OE line (FIG. 3C). The phenotypic results were repeatable and similar results were obtained with the GLK1 OE 2-6 line. The resistance to infection is consistent with the upregulation of defense related genes by GLK1.

Example 4

Global Analyses of Gene Expression

Global analyses of the effects of GLK1 over-expression (GLK1 OE) on gene expression were carried out by comparing three biological replicates of GLK OE against three biological replicates of the WT (wild-type) controls using the A. thaliana 25K Affymetrix^Rgene chip.
Total RNA samples obtained from biological replicates of three wild type, one GLK1 OE line 2-6 and two GLK1 OE line 5-7 leaves (Table 1), and TaGLK 1 OE (Table 2, see Example 6) Arabidopsis plants were used individually for the hybridizations with six Affymetrix Arabidopsis ATH1-121501 Gene Chips. The data was obtained from un-paired analyses of normalized values and represented differences that were statistically significant. The results identified 379 genes of which transcripts encoding one hundred and ninety-four genes were identified to be significantly upregulated and one hundred and forty-six downregulated more than 2 fold (Table 1; “Genes significantly up or down regulated 2-fold in Arabidopsis overexpressing (OE) Arabidopsis GLK1 (AtGLK1). Data analysed using FlexArray for normalization and T-Test”). The most highly accumulated transcripts encode proteins that have been identified in defense responses and include both PR genes as well as genes of enzymatic function implicated in biotic stress responses.
Table 2 identifies genes significantly up or down regulated 2-fold in Arabidopsis overexpressing (OE) wheat GLK1 (TaGLK1, see Example 6; Data analysed using FlexArray for normalization and T-Test)
Table 3 identifies common genes significantly up or down regulated 2-fold in Arabidopsis overexpressing (OE) Arabidopsis GLK1 (AtGLK1) and wheat GLK1 (TaGLK1; Data analysed using FlexArray for normalization and T-Test).
Affymetrix microarray analyses indicated that over-expression of wheat GLK1 (TaGLK1 Accession No. ABL10089) in Arabidopsis shows the regulation of similar genes as in AtGLK1 OE in Arabidopsis (Tables 1-3).
Some of the genes identified by the microarray were confirmed by RT-PCR. The most highly accumulated transcripts, confirmed by RT-PCR, include PB 1 (Phox bem1p domain protein) and ASN I (glutamine-dependent asparagine synthetase 1; FIG. 7). Phox-bem1p (PB1) and ASN1 transcripts are expressed in TaGLK1oe and AtGLK1oe Arabidopsis lines. This data validates the microarray data presented in Table 3 with the genes At1g77960 (PB1) and At3g47340 (ASN1). Taken together, these analyses indicate that TaGLK1 OE is capable of reprogramming identical networks of gene expression as that observed using AtGLK1 OE.
Increases in endogenous levels of ASN1 have been suggested to be associated with higher nitrogen status (Tian et al, 2007, J Biol Chem 282: 18532-18541). Over-expression of bacterial ASN1 is known to enhance the nitrogen status in Arabidopsis and soybean sink tissues (Lam et al, 2003, Plant Physiol 132: 926-935; Wan et al, 2006, Plant Biol (Stuttgart) 8: 271-276), enhanced vegetative growth (Giannino et al, 2007, Euphytica DOI 10.1007/s10681-007-9506-3) and increased synthesis of inulin (Sobolev et al, J Agric Food Chem 55, 10827-10831). The over-expression of AtGLK1 in maize cv. Hii2 produced transgenic maize that had darker green leaves when grown under midwinter (early December, Latitude 46N) greenhouse conditions suggestive of enhanced nitrogen utilization (FIG. 12). This demonstrating that over-expression of At and TaGLK1(s) presents an opportunity to modify nitrogen status in crop plants.
Additional highly accumulated transcripts represented potentially defense-related genes (FIG. 4). PR1, a gene indicative of the activation of the SAR (Uknes, S., et al., 1992, Plant Cell 4: 645-656) was downregulated as shown in both microarray and RT-PCR analyses (FIG. 4).
Microarray analyses of GLK2 over-expressing plants did not show the same profiles in defense related gene expression as observed in GLK1 over-expressing plants.

TABLE 1

Genes significantly up or down regulated 2-fold in Arabidopsis overexpressing
(OE) Arabidopsis GLK1 (AtGLK1). Data analysed using FlexArray for normalization and T-
Test

		AtGLK1
		OE Fold
Transcript ID	Gene_Title	change

At1g77960	Phox bem1p domain protein	42.25
At3g47340	glutamine-dependent asparagine synthetase 1 (ASN1)	12.29
At2g20570	golden2-like transcription factor (GLK1)	11.56
At2g39030	GCN5-related N-acetyltransferase (GNAT) family protein	10.94
At4g23680	major latex protein-related/MLP-related	9.92
At5g22500	acyl CoA reductase, putative/male-sterility protein, putative	8.42
At5g45670	GDSL-motif lipase/hydrolase family protein	8.01
At3g55970	oxidoreductase, 2OG-Fe(II) oxygenase family protein	8.01
At1g77970	hydroxyproline-rich glycoprotein family protein	7.85
At5g37300	expressed protein	7.39
At2g37770	aldo/keto reductase family protein	7.37
At1g35310	Bet v I allergen family protein	6.64
At3g56350	superoxide dismutase (Mn), putative	6.47
At2g20670	expressed protein	6.28
At5g44530	subtilase family protein	5.98
At3g60140	glycosyl hydrolase family 1 protein	5.92
At1g09350	galactinol synthase, putative	5.82
At1g13650	expressed protein	5.66
At3g11480	S-adenosyl-L-methionine:carboxyl methyltransferase family protein	5.60
At4g15210	beta-amylase (BMY1)/1,4-alpha-D-glucan maltohydrolase	5.40
At3g16460	jacalin lectin family protein	5.15
At1g73260	trypsin and protease inhibitor family protein/Kunitz family	4.95
At2g23000	serine carboxypeptidase S10 family protein	4.84
At2g21220	auxin-responsive protein, putative	4.73
At4g22470	protease inhibitor/seed storage/lipid transfer protein (LTP) family protein	4.69
At4g37990	mannitol dehydrogenase, putative (ELI3-2)	4.69
At1g56650	myb family transcription factor (MYB75)	4.67
At5g48490	protease inhibitor/seed storage/lipid transfer protein (LTP) family protein	4.49
At1g78170	expressed protein	4.47
At1g79460	ent-kaurene synthase/ent-kaurene synthetase B (KS) (GA2)	4.39
At2g35290	expressed protein	4.39
At1g14250	nucleoside phosphatase family protein/GDA1/CD39 family protein	4.37
At3g15720	glycoside hydrolase family 28 protein/polygalacturonase (pectinase)	4.31
	family protein
At5g24780	vegetative storage protein 2 (VSP2)	4.20
At5g62165	MADS-box protein (AGL42)	4.14
At1g09500	cinnamyl-alcohol dehydrogenase family/CAD family	4.14
At2g33380	calcium-binding RD20 protein (RD20)	4.10
At3g04000	short-chain dehydrogenase/reductase (SDR) family protein	3.99
At5g36910	thionin (THI2.2)	3.99
At1g52800	oxidoreductase, 2OG-Fe(II) oxygenase family protein	3.96
At2g46790	pseudo-response regulator, putative/timing of CAB expression 1-like	3.92
	protein,
At2g30790	photosystem II oxygen-evolving complex 23, putative	3.86
At5g09730	glycosyl hydrolase family 3 protein	3.83
At1g77210	sugar transporter, putative	3.79
At2g01890	purple acid phosphatase, putative	3.78
At5g42800	dihydroflavonol 4-reductase (dihydrokaempferol 4-reductase) (DFR)	3.74
At4g35060	heavy-metal-associated domain-containing protein/copper chaperone	3.73
	(CCH)-related
At5g49080	proline-rich extensin-like family protein	3.72
At5g35490	expressed protein (MRU1)	3.70
At1g10585	expressed protein	3.64
At5g23820	MD-2-related lipid recognition domain-containing protein/ML domain-	3.58
	containing protein
At1g35290	thioesterase family protein	3.55
At1g53885	senescence-associated protein-related	3.54
At2g47180	galactinol synthase, putative	3.49
At5g59480	haloacid dehalogenase-like hydrolase family protein	3.47
At4g09020	isoamylase, putative/starch debranching enzyme, putative	3.41
At5g50915	basic helix-loop-helix (bHLH) family protein	3.40
At2g15090	fatty acid elongase, putative	3.37
At2g42530	cold-responsive protein/cold-regulated protein (cor15b)	3.35
At1g80160	lactoylglutathione lyase family protein/glyoxalase I family protein	3.35
At2g37900	proton-dependent oligopeptide transport (POT) family protein	3.32
At5g59310	lipid transfer protein 4 (LTP4)	3.25
At1g17020	oxidoreductase, 2OG-Fe(II) oxygenase family protein	3.19
At2g39920	acid phosphatase class B family protein	3.18
At4g37770	1-aminocyclopropane-1-carboxylate synthase, putative/ACC synthase,	3.13
	putative
At3g15950	DNA topoisomerase-related	3.11
At3g08920	rhodanese-like domain-containing protein	3.05
At2g34810	FAD-binding domain-containing protein	3.04
At1g49430	long-chain-fatty-acid--CoA ligase/long-chain acyl-CoA synthetase	3.03
At3g09260	glycosyl hydrolase family 1 protein	2.97
At2g38870	protease inhibitor, putative	2.96
At5g50950	fumarate hydratase, putative/fumarase, putative	2.94
At1g77580	myosin heavy chain-related	2.92
At3g28220	meprin and TRAF homology domain-containing protein/MATH domain-	2.92
	containing protein
At1g34040	alliinase family protein /// alliinase family protein	2.92
At2g29490	glutathione S-transferase, putative	2.91
At1g52690	late embryogenesis abundant protein, putative/LEA protein, putative	2.88
At4g12500	protease inhibitor/seed storage/lipid transfer protein (LTP) family protein	2.86
At2g46680	homeobox-leucine zipper protein 7 (HB-7)/HD-ZIP transcription factor 7	2.85
At2g21650	myb family transcription factor	2.81
At2g38180	GDSL-motif lipase/hydrolase family protein	2.81
At3g28740	cytochrome P450 family protein	2.81
At2g48020	sugar transporter, putative	2.81
At1g73870	zinc finger (B-box type) family protein	2.77
At4g12490	protease inhibitor/seed storage/lipid transfer protein (LTP) family protein	2.77
At3g13672	seven in absentia (SINA) family protein	2.74
At1g18870	isochorismate synthase, putative/isochorismate mutase, putative	2.73
At2g42190	expressed protein	2.72
At5g67520	adenylylsulfate kinase, putative	2.70
At5g24420	glucosamine/galactosamine-6-phosphate isomerase-related	2.69
At2g43510	trypsin inhibitor, putative	2.69
At1g56170	transcription factor, putative	2.67
At1g16530	LOB domain protein 3/lateral organ boundaries domain protein 3	2.67
	(LBD3)
At2g22240	inositol-3-phosphate synthase isozyme 2/myo-inositol-1-phosphate	2.66
	synthase 2/MI-1-P synthase 2/IPS 2
At4g21910	MATE efflux family protein	2.66
At5g24160	squalene monooxygenase 1,2/squalene epoxidase 1,2 (SQP1,2)	2.65
At3g29810	phytochelatin synthetase family protein/COBRA cell expansion protein	2.64
	COBL2
At1g58290	glutamyl-tRNA reductase 1/GluTR (HEMA1)	2.64
At1g64200	vacuolar ATP synthase subunit E, putative	2.64
At4g21680	proton-dependent oligopeptide transport (POT) family protein	2.63
At4g10850	nodulin MtN3 family protein	2.62
At1g69160	expressed protein	2.60
At2g36050	ovate protein-related	2.60
At5g49360	glycosyl hydrolase family 3 protein	2.60
At5g47610	zinc finger (C3HC4-type RING finger) family protein	2.60
At3g20100	cytochrome P450 family protein	2.60
At1g15010	expressed protein	2.60
At5g05600	oxidoreductase, 2OG-Fe(II) oxygenase family protein	2.59
At4g19020	chromomethylase 2 (CMT2)	2.57
At2g34070	expressed protein	2.56
At5g43580	protease inhibitor, putative	2.56
At5g05860	UDP-glucoronosyl/UDP-glucosyl transferase family protein	2.56
At4g13560	late embryogenesis abundant domain-containing protein/LEA domain-	2.55
	containing protein
At1g55960	expressed protein	2.54
At4g23600	coronatine-responsive tyrosine aminotransferase/tyrosine transaminase	2.53
At2g36080	DNA-binding protein, putative	2.52
At5g24150	squalene monooxygenase 1,1/squalene epoxidase 1,1 (SQP1,1)	2.52
At1g79160	expressed protein	2.52
At1g02220	no apical meristem (NAM) family protein	2.50
At4g39510	cytochrome P450 family protein	2.50
At3g29575	expressed protein	2.50
At5g65020	annexin 2 (ANN2)	2.49
At2g30540	glutaredoxin family protein	2.48
At5g52570	beta-carotene hydroxylase, putative	2.47
At3g02480	ABA-responsive protein-related	2.46
At5g02940	expressed protein	2.45
At2g02990	ribonuclease 1 (RNS1)	2.45
At5g59220	protein phosphatase 2C, putative/PP2C, putative	2.45
At1g66760	MATE efflux family protein	2.44
At5g17220	glutathione S-transferase, putative	2.44
At4g27440	protochlorophyllide reductase B, chloroplast/PCR B/NADPH-	2.44
	protochlorophyllide oxidoreductase B (PORB)
At4g01430	nodulin MtN21 family protein	2.44
At2g39330	jacalin lectin family protein	2.44
At1g11300	S-locus lectin protein kinase family protein	2.44
At1g62660	beta-fructosidase (BFRUCT3)/beta-fructofuranosidase/invertase,	2.43
	vacuolar
At1g59930	hypothetical protein	2.43
At4g17470	palmitoyl protein thioesterase family protein	2.43
At3g02380	zinc finger protein CONSTANS-LIKE 2 (COL2)	2.43
At5g01520	zinc finger (C3HC4-type RING finger) family protein	2.43
At4g33666	expressed protein	2.42
At2g25900	zinc finger (CCCH-type) family protein	2.42
At1g68190	zinc finger (B-box type) family protein	2.41
At4g33150	lysine-ketoglutarate reductase/saccharopine dehydrogenase bifunctional	2.41
	enzyme
At3g14770	nodulin MtN3 family protein	2.40
At1g02340	long hypocotyl in far-red 1 (HFR1)/reduced phytochrome signaling	2.40
	(REP1)/basic helix-loop-helix FBI1 protein (FBI1)/reduced sensitivity
	to far-red light (RSF1)/bHLH protein 26 (BHLH026) (BHLH26)
At5g44050	MATE efflux family protein	2.39
At4g19430	expressed protein	2.39
At3g06500	beta-fructofuranosidase, putative/invertase, putative/saccharase,	2.39
	putative/beta-fructosidase, putative
At5g57340	expressed protein	2.38
At1g20190	expansin, putative (EXP11)	2.38
At4g37530	peroxidase 50 (PER50) (P50) (PRXR2)	2.37
At1g10370	glutathione S-transferase, putative (ERD9)	2.37
At4g22870	leucoanthocyanidin dioxygenase, putative/anthocyanidin synthase,	2.36
	putative
At2g03600	expressed protein	2.36
At5g35510	hypothetical protein	2.36
At2g39420	esterase/lipase/thioesterase family protein	2.35
At1g19640	S-adenosyl-L-methionine:jasmonic acid carboxyl methyltransferase	2.35
	(JMT)
At4g16730	terpene synthase/cyclase family protein	2.35
At1g44350	IAA-amino acid hydrolase 6, putative (ILL6)/IAA-Ala hydrolase, putative	2.34
At1g71140	MATE efflux family protein	2.33
At1g52890	no apical meristem (NAM) family protein	2.33
At2g16660	nodulin family protein	2.31
At4g12480	protease inhibitor/seed storage/lipid transfer protein (LTP) family protein	2.30
At1g26920	expressed protein	2.29
At1g60030	xanthine/uracil permease family protein	2.29
At1g55020	lipoxygenase (LOX1)	2.28
At5g04490	phosphatidate cytidylyltransferase family protein	2.28
At5g35480	expressed protein	2.27
At3g56080	dehydration-responsive protein-related	2.27
At2g46640	hypothetical protein	2.26
At2g40880	cysteine protease inhibitor, putative/cystatin, putative (FL3-27)	2.26
At2g43535	trypsin inhibitor, putative	2.25
At1g22550	proton-dependent oligopeptide transport (POT) family protein	2.22
At2g36590	proline transporter, putative	2.22
At2g24580	sarcosine oxidase family protein	2.22
At4g16590	glucosyltransferase-related	2.22
At2g15960	expressed protein	2.21
At1g13080	cytochrome P450 family protein	2.20
At5g45960	GDSL-motif lipase/hydrolase family protein	2.20
At3g55110	ABC transporter family protein	2.20
At3g57680	peptidase S41 family protein	2.19
At5g15960	stress-responsive protein (KIN1)	2.19
At4g08870	arginase, putative /// arginase, putative	2.19
At5g65390	arabinogalactan-protein (AGP7)	2.17
At4g11650	osmotin-like protein (OSM34)	2.17
At2g22470	arabinogalactan-protein (AGP2)	2.17
At1g56580	expressed protein	2.17
At4g17500	ethylene-responsive element-binding protein 1 (ERF1)/EREBP-2	2.17
	protein
At3g62950	glutaredoxin family protein	2.16
At4g01080	expressed protein	2.16
At1g75580	auxin-responsive protein, putative	2.16
At1g76090	S-adenosyl-methionine-sterol-C-methyltransferase	2.15
At1g69490	no apical meristem (NAM) family protein	2.15
At4g15660	glutaredoxin family protein	2.14
At3g29320	glucan phosphorylase, putative	2.13
At5g54060	glycosyltransferase family protein	2.13
At4g37800	xyloglucan:xyloglucosyl transferase, putative/xyloglucan	2.12
	endotransglycosylase, putative/endo-xyloglucan transferase, putative
At1g03020	glutaredoxin family protein	2.12
At1g75450	FAD-binding domain-containing protein/cytokinin oxidase family protein	2.11
At3g46660	UDP-glucoronosyl/UDP-glucosyl transferase family protein	2.11
At1g70820	phosphoglucomutase, putative/glucose phosphomutase, putative	2.11
At1g17970	zinc finger (C3HC4-type RING finger) family protein	2.10
At4g16690	esterase/lipase/thioesterase family protein	2.10
At5g06870	polygalacturonase inhibiting protein 2 (PGIP2)	2.10
At4g27410	no apical meristem (NAM) family protein (RD26)	2.09
At3g01420	pathogen-responsive alpha-dioxygenase, putative	2.09
At1g05340	expressed protein	2.09
At1g29395	stress-responsive protein, putative	2.09
At3g11900	amino acid transporter family protein	2.09
At4g21620	glycine-rich protein	2.08
At4g18440	adenylosuccinate lyase, putative/adenylosuccinase, putative	2.08
At1g20350	mitochondrial import inner membrane translocase subunit Tim17,	2.08
	putative
At1g74010	strictosidine synthase family protein	2.08
At3g61060	F-box family protein/lectin-related	2.07
At3g07350	expressed protein	2.07
At5g13400	proton-dependent oligopeptide transport (POT) family protein	2.07
At1g06570	4-hydroxyphenylpyruvate dioxygenase (HPD)	2.05
At1g76790	O-methyltransferase family 2 protein	2.05
At1g71080	expressed protein	2.05
At5g57550	xyloglucan:xyloglucosyl transferase/xyloglucan endotransglycosylase/	2.04
	endo-xyloglucan transferase (XTR3)
At2g37180	plasma membrane intrinsic protein 2B (PIP2B)/aquaporin PIP2.2	2.04
	(PIP2.2)
At4g24380	expressed protein	2.04
At2g28190	superoxide dismutase (Cu—Zn), chloroplast (SODCP)/copper/zinc	2.03
	superoxide dismutase (CSD2)
At5g45840	leucine-rich repeat transmembrane protein kinase, putative	2.03
At2g37760	aldo/keto reductase family protein	2.03
At2g26690	nitrate transporter (NTP2)	2.02
At2g36870	xyloglucan:xyloglucosyl transferase, putative/xyloglucan	2.02
	endotransglycosylase, putative/endo-xyloglucan transferase, putative
At5g48180	kelch repeat-containing protein	2.02
At5g54160	quercetin 3-O-methyltransferase 1/flavonol 3-O-methyltransferase 1/	2.02
	caffeic acid/5-hydroxyferulic acid O-methyltransferase (OMT1)
At1g60190	armadillo/beta-catenin repeat family protein/U-box domain-containing	2.02
	protein
At4g14130	xyloglucan:xyloglucosyl transferase, putative/xyloglucan	2.02
	endotransglycosylase, putative/endo-xyloglucan transferase, putative
	(XTR7)
At3g14060	expressed protein /// expressed protein	2.01
At3g23410	alcohol oxidase-related	2.01
At1g54040	kelch repeat-containing protein	2.01
At1g12520	superoxide dismutase copper chaperone, putative	2.01
At3g05180	GDSL-motif lipase/hydrolase family protein	2.01
At5g65870	phytosulfokines 5 (PSK5)	2.01
At3g50770	calmodulin-related protein, putative	2.00
At5g02580	expressed protein	2.00
At5g24120	RNA polymerase sigma subunit SigE (sigE)/sigma-like factor (SIG5)	2.00
At5g13220	expressed protein	2.00
At1g56670	GDSL-motif lipase/hydrolase family protein	2.00
At1g24150	formin homology 2 domain-containing protein/FH2 domain-containing	−2.02
	protein
At3g05730	expressed protein	−2.02
At4g24190	shepherd protein (SHD)/clavata formation protein, putative	−2.03
At4g27300	S-locus protein kinase, putative	−2.03
At5g07030	aspartyl protease family protein	−2.04
At1g27030	expressed protein	−2.04
At5g04950	nicotianamine synthase, putative	−2.04
At5g11670	malate oxidoreductase, putative	−2.05
At2g31060	elongation factor family protein	−2.05
At2g33210	chaperonin, putative	−2.06
At5g54610	ankyrin repeat family protein	−2.06
At2g36970	UDP-glucoronosyl/UDP-glucosyl transferase family protein	−2.06
At3g57260	glycosyl hydrolase family 17 protein	−2.07
At4g16870	—	−2.07
At2g22500	mitochondrial substrate carrier family protein	−2.07
At1g35230	arabinogalactan-protein (AGP5)	−2.07
At2g41180	sigA-binding protein-related	−2.07
At2g16060	non-symbiotic hemoglobin 1 (HB1) (GLB1)	−2.08
At3g15400	anther development protein, putative	−2.08
At1g03210	phenazine biosynthesis PhzC/PhzF family protein	−2.08
At3g09940	monodehydroascorbate reductase, putative	−2.08
At2g24850	aminotransferase, putative	−2.10
At5g60950	phytochelatin synthetase-related	−2.11
At1g76960	expressed protein	−2.11
At1g48210	serine/threonine protein kinase, putative	−2.11
At2g02130	plant defensin-fusion protein, putative (PDF2.3)	−2.12
At4g23130	receptor-like protein kinase 6 (RLK6)	−2.12
At2g26020	plant defensin-fusion protein, putative (PDF1.2b)	−2.12
At1g56120	leucine-rich repeat family protein/protein kinase family protein	−2.14
At1g08050	zinc finger (C3HC4-type RING finger) family protein	−2.15
At4g18250	receptor serine/threonine kinase, putative	−2.16
At2g38940	phosphate transporter (PT2)	−2.16
At1g01340	cyclic nucleotide-regulated ion channel (CNGC10) (ACBK1)	−2.17
At3g05650	disease resistance family protein	−2.17
At5g08260	serine carboxypeptidase S10 family protein	−2.17
At1g08450	calreticulin 3 (CRT3)	−2.18
At1g57630	disease resistance protein (TIR class), putative	−2.18
At1g19960	expressed protein	−2.18
At4g14400	ankyrin repeat family protein	−2.19
At3g02040	glycerophosphoryl diester phosphodiesterase family protein	−2.20
At5g62210	embryo-specific protein-related	−2.21
At3g24900	disease resistance family protein/LRR family protein	−2.21
At2g14610	pathogenesis-related protein 1 (PR-1)	−2.22
At5g54720	ankyrin repeat family protein	−2.23
At3g50480	broad-spectrum mildew resistance RPW8 family protein	−2.23
At5g52810	ornithine cyclodeaminase/mu-crystallin family protein	−2.23
At2g34260	transducin family protein/WD-40 repeat family protein	−2.24
At2g22170	lipid-associated family protein	−2.24
At3g53980	protease inhibitor/seed storage/lipid transfer protein (LTP) family protein	−2.25
At5g25260	expressed protein	−2.25
At4g20780	calcium-binding protein, putative	−2.25
At5g44420	plant defensin protein, putative (PDF1.2a)	−2.26
At4g21830	methionine sulfoxide reductase domain-containing protein/SelR	−2.26
	domain-containing protein
At2g20870	cell wall protein precursor, putative	−2.26
At4g23200	protein kinase family protein	−2.27
At3g26210	cytochrome P450 71B23, putative (CYP71B23)	−2.27
At5g39670	calcium-binding EF hand family protein	−2.27
At3g60420	expressed protein	−2.27
At2g40750	WRKY family transcription factor	−2.28
At5g02760	protein phosphatase 2C family protein/PP2C family protein	−2.28
At3g44750	histone deacetylase, putative (HD2A)	−2.30
At1g80270	DNA-binding protein, putative	−2.30
At1g10340	ankyrin repeat family protein	−2.31
At1g24020	Bet v I allergen family protein	−2.31
At5g20400	oxidoreductase, 2OG-Fe(II) oxygenase family protein	−2.32
At3g51330	aspartyl protease family protein	−2.32
At5g58940	protein kinase family protein	−2.34
At1g35710	leucine-rich repeat transmembrane protein kinase, putative	−2.34
At5g45380	sodium:solute symporter family protein	−2.35
At4g11850	phospholipase D gamma 1/PLD gamma 1 (PLDGAMMA1)	−2.37
At4g00700	C2 domain-containing protein	−2.39
At5g52310	low-temperature-responsive protein 78 (LTI78)/desiccation-responsive	−2.39
	protein 29A (RD29A)
At5g22580	expressed protein	−2.40
At3g05660	disease resistance family protein	−2.40
At5g12940	leucine-rich repeat family protein	−2.42
At4g34135	UDP-glucoronosyl/UDP-glucosyl transferase family protein	−2.42
At1g52200	expressed protein	−2.42
At4g23220	protein kinase family protein	−2.46
At2g04450	MutT/nudix family protein	−2.47
At1g67990	caffeoyl-CoA 3-O-methyltransferase, putative	−2.50
At4g23150	protein kinase family protein	−2.50
At1g21250	wall-associated kinase 1 (WAK1)	−2.52
At5g28540	luminal binding protein 1 (BiP-1) (BP1)	−2.53
At3g09020	alpha 1,4-glycosyltransferase family protein/glycosyltransferase sugar-	−2.54
	binding DXD motif-containing protein
At2g02930	glutathione S-transferase, putative	−2.55
At5g61160	transferase family protein	−2.55
At2g02810	UDP-galactose/UDP-glucose transporter	−2.56
At3g48640	expressed protein	−2.58
At4g21380	S-locus protein kinase, putative (ARK3)	−2.58
At4g14365	zinc finger (C3HC4-type RING finger) family protein/ankyrin repeat	−2.61
	family protein
At3g01290	band 7 family protein	−2.63
At1g30900	vacuolar sorting receptor, putative	−2.64
At2g26440	pectinesterase family protein	−2.64
At4g02380	late embryogenesis abundant 3 family protein/LEA3 family protein	−2.67
At1g13470	expressed protein	−2.71
At3g23110	disease resistance family protein /// disease resistance family protein	−2.73
At5g64510	expressed protein	−2.74
At5g59670	leucine-rich repeat protein kinase, putative	−2.75
At1g75040	pathogenesis-related protein 5 (PR-5)	−2.76
At5g42900	expressed protein	−2.77
At1g66850	protease inhibitor/seed storage/lipid transfer protein (LTP) family protein	−2.78
At4g01700	chitinase, putative	−2.79
At1g77510	protein disulfide isomerase, putative	−2.82
At5g07550	glycine-rich protein (GRP19)	−2.84
At1g31690	copper amine oxidase, putative	−2.87
At3g25010	disease resistance family protein	−2.90
At1g75940	glycosyl hydrolase family 1 protein/anther-specific protein ATA27	−2.93
At1g04980	thioredoxin family protein	−2.94
At4g23140	receptor-like protein kinase 5 (RLK5)	−2.95
At2g47130	short-chain dehydrogenase/reductase (SDR) family protein	−2.97
At5g20630	germin-like protein (GER3)	−3.01
At4g11890	protein kinase family protein	−3.03
At1g68875	expressed protein	−3.04
At4g10500	oxidoreductase, 2OG-Fe(II) oxygenase family protein	−3.17
At5g52760	heavy-metal-associated domain-containing protein	−3.20
At3g48080	lipase class 3 family protein/disease resistance protein-related	−3.31
At5g52740	heavy-metal-associated domain-containing protein	−3.31
At5g59680	leucine-rich repeat protein kinase, putative	−3.34
At4g17030	expansin-related	−3.35
At3g16530	legume lectin family protein	−3.57
At4g03450	ankyrin repeat family protein	−3.60
At2g26560	patatin, putative	−3.60
At3g23550	MATE efflux family protein	−3.61
At1g14870	expressed protein	−3.73
At1g67810	Fe—S metabolism associated domain-containing protein	−3.77
At3g11010	disease resistance family protein/LRR family protein	−3.82
At3g16670	expressed protein	−3.86
At5g55450	protease inhibitor/seed storage/lipid transfer protein (LTP) family protein	−3.90
At5g20790	expressed protein	−3.93
At2g14560	expressed protein	−3.96
At1g09080	luminal binding protein 3 (BiP-3) (BP3)	−4.12
At4g25110	latex-abundant family protein (AMC2)/caspase family protein	−4.21
At4g08300	nodulin MtN21 family protein	−4.23
At5g18470	curculin-like (mannose-binding) lectin family protein	−4.23
At3g51590	lipid transfer protein, putative	−5.14
At1g29720	protein kinase family protein	−5.41
At3g57240	beta-1,3-glucanase (BG3)	−5.50
At5g23020	2-isopropylmalate synthase 2 (IMS2)	−8.88

TABLE 2

Genes significantly up or down regulated 2-fold in Arabidopsis overexpressing
(OE) wheat GLK1 (TaGLK1). Data analysed using FlexArray for normalization and T-Test

		TaGLK1
		OE Fold
Transcript ID	Gene Title	change

At3g08770	lipid transfer protein 6 (LTP6)	74.83
At2g33850	expressed protein	63.70
At1g72260	thionin (THI2.1)	47.99
At2g42840	protodermal factor 1 (PDF1)	47.96
At3g09260	glycosyl hydrolase family 1 protein	26.10
At4g37410	cytochrome P450, putative	25.94
At1g62500	protease inhibitor/seed storage/lipid transfer protein (LTP) family protein	25.44
At5g45670	GDSL-motif lipase/hydrolase family protein	24.42
At4g22490	protease inhibitor/seed storage/lipid transfer protein (LTP) family protein	21.94
At4g29030	glycine-rich protein	19.68
At1g77960	Phox-bem1p domain protein	18.56
At2g28790	osmotin-like protein, putative	16.93
At5g36910	thionin (THI2.2)	16.22
At2g20870	cell wall protein precursor, putative	15.83
At1g54010	myrosinase-associated protein, putative	15.49
At1g28290	pollen Ole e 1 allergen and extensin family protein	15.44
At1g54020	myrosinase-associated protein, putative	14.89
At5g23940	transferase family protein	14.76
At5g33370	GDSL-motif lipase/hydrolase family protein	14.45
At3g15950	DNA topoisomerase-related	12.88
At5g08000	glycosyl hydrolase family protein 17	12.68
At5g42800	dihydroflavonol 4-reductase (dihydrokaempferol 4-reductase) (DFR)	12.64
At3g16430	jacalin lectin family protein	12.56
At4g29700	type I phosphodiesterase/nucleotide pyrophosphatase family protein	11.56
At5g24420	glucosamine/galactosamine-6-phosphate isomerase-related	11.34
At2g39310	jacalin lectin family protein	10.72
At4g19430	expressed protein	10.34
At1g24020	Bet v I allergen family protein	10.18
At4g22870	leucoanthocyanidin dioxygenase, putative/anthocyanidin synthase,	10.13
	putative
At3g02550	LOB domain protein 41/lateral organ boundaries domain protein 41	10.00
	(LBD41)
At4g17470	palmitoyl protein thioesterase family protein	9.67
At4g23680	major latex protein-related/MLP-related	8.76
At5g20740	invertase/pectin methylesterase inhibitor family protein	8.74
At1g18250	thaumatin, putative	8.70
At2g14900	gibberellin-regulated family protein	8.65
At4g11310	cysteine proteinase, putative	8.65
At1g76790	O-methyltransferase family 2 protein	8.45
At5g07030	aspartyl protease family protein	8.32
At1g08560	syntaxin-related protein KNOLLE (KN)/syntaxin 111 (SYP111)	8.12
At5g23820	MD-2-related lipid recognition domain-containing protein/ML domain-	8.06
	containing protein
At5g28640	SSXT protein-related/glycine-rich protein	7.97
At4g23800	high mobility group (HMG1/2) family protein	7.95
At5g47500	pectinesterase family protein	7.92
At4g33260	WD-40 repeat family protein	7.86
At2g21140	hydroxyproline-rich glycoprotein family protein	7.85
At3g02640	expressed protein	7.83
At5g16250	expressed protein	7.75
At1g02730	cellulose synthase family protein	7.68
At1g31710	copper amine oxidase, putative	7.55
At1g10640	polygalacturonase, putative/pectinase, putative	7.55
At3g55970	oxidoreductase, 2OG-Fe(II) oxygenase family protein	7.33
At5g55180	glycosyl hydrolase family 17 protein	7.27
At5g17220	glutathione S-transferase, putative	7.26
At2g38530	nonspecific lipid transfer protein 2 (LTP2)	7.21
At4g15910	drought-responsive protein/drought-induced protein (Di21)	7.11
At4g02290	glycosyl hydrolase family 9 protein	7.07
At3g22880	meiotic recombination protein, putative	6.86
At5g54060	glycosyltransferase family protein	6.85
At2g43520	trypsin inhibitor, putative	6.81
At4g31840	plastocyanin-like domain-containing protein	6.81
At3g17680	expressed protein	6.68
At3g48350	cysteine proteinase, putative	6.66
At4g12490	protease inhibitor/seed storage/lipid transfer protein (LTP) family protein	6.61
At4g15830	expressed protein	6.58
At3g16450	jacalin lectin family protein	6.54
At1g18370	kinesin motor family protein (NACK1)	6.50
At2g02850	plastocyanin-like domain-containing protein/plantacyanin, putative	6.38
At1g03495	pseudogene, transferase family	6.17
At1g06100	fatty acid desaturase family protein	6.13
At4g12500	protease inhibitor/seed storage/lipid transfer protein (LTP) family protein	6.13
At4g34160	cyclin delta-3 (CYCD3)	6.11
At2g38870	protease inhibitor, putative	6.07
At2g23000	serine carboxypeptidase S10 family protein	6.04
At3g05600	epoxide hydrolase, putative	6.01
At1g19610	plant defensin-fusion protein, putative (PDF1.4)	5.92
At3g16390	jacalin lectin family protein	5.90
At5g51750	subtilase family protein	5.87
At1g13710	cytochrome P450 family protein	5.72
At1g70710	endo-1,4-beta-glucanase (EGASE)/cellulase	5.71
At1g43800	acyl-(acyl-carrier-protein) desaturase, putative/stearoyl-ACP	5.67
	desaturase, putative
At3g15720	glycoside hydrolase family 28 protein/polygalacturonase (pectinase)	5.67
At3g46640	myb family transcription factor	5.63
At4g29020	glycine-rich protein	5.56
At3g21770	peroxidase 30 (PER30) (P30) (PRXR9)	5.51
At5g04530	beta-ketoacyl-CoA synthase family protein	5.46
At3g02120	hydroxyproline-rich glycoprotein family protein	5.45
At1g80080	leucine-rich repeat family protein	5.44
At4g32830	protein kinase, putative	5.41
At2g25880	serine/threonine protein kinase, putative	5.40
At5g17700	MATE efflux family protein	5.40
At4g02850	phenazine biosynthesis PhzC/PhzF family protein	5.39
At1g52040	myrosinase-binding protein, putative (F-ATMBP)	5.37
At4g02800	expressed protein	5.34
At1g14900	high-mobility-group protein/HMG-I/Y protein	5.33
At5g43020	leucine-rich repeat transmembrane protein kinase, putative	5.31
At5g22430	expressed protein	5.26
At1g75240	zinc finger homeobox family protein/ZF-HD homeobox family protein	5.26
At2g34490	cytochrome P450 family protein	5.25
At5g37300	expressed protein	5.24
At3g47340	glutamine-dependent asparagine synthetase 1 (ASN1)	5.24
At4g05520	calcium-binding EF hand family protein	5.21
At2g28210	carbonic anhydrase family protein	5.21
At3g50070	cyclin family protein	5.19
At1g44110	cyclin, putative	5.12
At3g16470	jacalin lectin family protein	5.10
At2g21640	expressed protein	5.10
At5g01600	ferritin 1 (FER1)	5.06
At2g43800	formin homology 2 domain-containing protein/FH2 domain-containing	5.04
	protein
At3g18000	phosphoethanolamine N-methyltransferase 1/PEAMT 1 (NMT1)	5.03
At5g13840	WD-40 repeat family protein	5.01
At1g14440	zinc finger homeobox family protein/ZF-HD homeobox family protein	5.00
At3g28290	integrin-related protein 14a	4.98
At3g04290	GDSL-motif lipase/hydrolase family protein	4.89
At4g02360	expressed protein	4.86
At5g25090	plastocyanin-like domain-containing protein	4.82
At3g25980	mitotic spindle checkpoint protein, putative (MAD2)	4.80
At2g38750	annexin 4 (ANN4)	4.78
At2g42610	expressed protein	4.78
At3g28500	60S acidic ribosomal protein P2 (RPP2C)	4.77
At3g28740	cytochrome P450 family protein	4.76
At1g01600	cytochrome P450, putative	4.74
At3g53190	pectate lyase family protein	4.74
At4g14090	UDP-glucoronosyl/UDP-glucosyl transferase family protein	4.73
At3g23890	DNA topoisomerase, ATP-hydrolyzing/DNA topoisomerase II/DNA	4.71
	gyrase (TOP2)
At1g22770	gigantea protein (GI)	4.70
At1g64200	vacuolar ATP synthase subunit E, putative	4.68
At5g54510	auxin-responsive GH3 protein, putative (DFL-1)	4.61
At5g36710	expressed protein	4.60
At1g04110	subtilase family protein	4.56
At1g54050	17.4 kDa class III heat shock protein (HSP17.4-CIII)	4.56
At4g37750	ovule development protein aintegumenta (ANT)	4.56
At3g55110	ABC transporter family protein	4.53
At1g24070	glycosyl transferase family 2 protein	4.52
At5g38120	4-coumarate--CoA ligase family protein/4-coumaroyl-CoA synthase	4.50
	family protein
At4g12480	protease inhibitor/seed storage/lipid transfer protein (LTP) family protein	4.45
At5g01870	lipid transfer protein, putative	4.43
At2g25060	plastocyanin-like domain-containing protein	4.43
At4g39510	cytochrome P450 family protein	4.42
At4g14770	tesmin/TSO1-like CXC domain-containing protein	4.42
At4g12470	protease inhibitor/seed storage/lipid transfer protein (LTP) family protein	4.38
At5g48485	protease inhibitor/seed storage/lipid transfer protein (LTP) family protein	4.35
At1g07050	CONSTANS-like protein-related	4.32
At1g73620	thaumatin-like protein, putative/pathogenesis-related protein, putative	4.30
At5g57785	expressed protein	4.30
At1g79840	homeobox-leucine zipper protein 10 (HB-10)/HD-ZIP transcription	4.29
	factor 10/homeobox protein (GLABRA2)
At2g19590	1-aminocyclopropane-1-carboxylate oxidase, putative/ACC oxidase,	4.28
	putative
At1g52410	caldesmon-related	4.26
At5g44560	SNF7 family protein	4.26
At2g21650	myb family transcription factor	4.24
At5g13220	expressed protein	4.23
At2g02100	plant defensin-fusion protein, putative (PDF2.2)	4.23
At1g76540	cell division control protein, putative	4.18
At5g64080	protease inhibitor/seed storage/lipid transfer protein (LTP) family protein	4.17
At1g62360	homeobox protein SHOOT MERISTEMLESS (STM)	4.17
At1g12845	expressed protein	4.17
At3g51280	male sterility MS5, putative	4.17
At3g14190	expressed protein	4.16
At1g48330	expressed protein	4.15
At5g19110	extracellular dermal glycoprotein-related/EDGP-related	4.08
At2g39330	jacalin lectin family protein	4.08
At1g50490	ubiquitin-conjugating enzyme 20 (UBC20)	4.04
At4g27860	integral membrane family protein	4.04
At1g25450	very-long-chain fatty acid condensing enzyme, putative	4.03
At4g30140	GDSL-motif lipase/hydrolase family protein	3.99
At2g23170	auxin-responsive GH3 family protein	3.99
At1g67750	pectate lyase family protein	3.96
At1g72970	glucose-methanol-choline (GMC) oxidoreductase family protein	3.95
At1g70830	Bet v I allergen family protein	3.95
At3g13175	expressed protein	3.95
At4g34250	fatty acid elongase, putative	3.93
At1g51460	ABC transporter family protein	3.93
At2g38760	annexin 3 (ANN3)	3.92
At5g15230	gibberellin-regulated protein 4 (GASA4)	3.91
At3g16460	jacalin lectin family protein	3.91
At2g43620	chitinase, putative	3.90
At1g56650	myb family transcription factor (MYB75)	3.90
At3g11600	expressed protein	3.89
At2g40300	ferritin, putative	3.89
At2g22840	expressed protein	3.89
At3g23730	xyloglucan:xyloglucosyl transferase, putative	3.88
At4g03210	xyloglucan:xyloglucosyl transferase, putative	3.87
At1g66370	myb family transcription factor (MYB113)	3.86
At1g62480	vacuolar calcium-binding protein-related	3.85
At3g29590	transferase family protein	3.85
At4g11650	osmotin-like protein (OSM34)	3.85
At1g20030	pathogenesis-related thaumatin family protein	3.84
At1g06350	fatty acid desaturase family protein	3.83
At4g11190	disease resistance-responsive family protein/dirigent family protein	3.80
At3g52110	expressed protein	3.80
At3g01140	myb family transcription factor (MYB106)	3.79
At5g58390	peroxidase, putative	3.79
At3g06030	NPK1-related protein kinase, putative (ANP3)	3.79
At1g55260	protease inhibitor/seed storage/lipid transfer protein (LTP) family protein	3.76
At1g10522	expressed protein	3.75
At3g17990	phosphoethanolamine N-methyltransferase 1/PEAMT 1 (NMT1)	3.75
At5g60890	receptor-like protein kinase (ATR1) (MYB34)	3.75
At4g00480	myc-related transcription factor (MYC1)	3.72
At1g50240	armadillo/beta-catenin repeat family protein	3.72
At2g39030	GCN5-related N-acetyltransferase (GNAT) family protein	3.70
At1g17860	trypsin and protease inhibitor family protein/Kunitz family	3.70
At4g38660	thaumatin, putative	3.69
At3g23830	glycine-rich RNA-binding protein, putative	3.69
At3g21300	RNA methyltransferase family protein	3.66
At2g37640	expansin, putative (EXP3)	3.64
At5g17160	expressed protein	3.64
At2g42570	expressed protein	3.63
At4g03100	rac GTPase activating protein, putative	3.62
At5g16190	glycosyl transferase family 2 protein	3.60
At1g53885	senescence-associated protein-related	3.60
At4g37740	expressed protein	3.59
At1g78490	cytochrome P450 family protein	3.58
At1g17380	expressed protein	3.58
At1g73260	trypsin and protease inhibitor family protein/Kunitz family protein	3.57
At1g74430	myb family transcription factor (MYB95)	3.54
At5g67270	microtubule-associated EB1 family protein	3.53
At4g31290	ChaC-like family protein	3.52
At5g23310	superoxide dismutase (Fe)/iron superoxide dismutase 3 (FSD3)	3.51
At1g75780	tubulin beta-1 chain (TUB1)	3.50
At1g33170	dehydration-responsive family protein	3.49
At3g56070	peptidyl-prolyl cis-trans isomerase, putative	3.45
At5g60100	pseudo-response regulator 3 (APRR3)	3.44
At1g03780	targeting protein-related	3.43
At3g44990	xyloglucan:xyloglucosyl transferase, putative/xyloglucan	3.41
	endotransglycosylase, putative/endo-xyloglucan transferase, putative
At5g45700	NLI interacting factor (NIF) family protein	3.41
At2g34810	FAD-binding domain-containing protein	3.41
At1g69200	pfkB-type carbohydrate kinase family protein	3.39
At1g50010	tubulin alpha-2/alpha-4 chain (TUA2)	3.39
At5g48360	formin homology 2 domain-containing protein/FH2 domain protein	3.38
At5g37010	expressed protein	3.38
At3g48500	expressed protein	3.37
At3g12500	basic endochitinase	3.37
At3g54820	aquaporin, putative	3.37
At5g39790	5′-AMP-activated protein kinase beta-1 subunit-related	3.34
At3g27690	chlorophyll A-B binding protein (LHCB2:4)	3.33
At3g26330	cytochrome P450 family protein	3.32
At5g65020	annexin 2 (ANN2)	3.32
At1g02690	importin alpha-2 subunit, putative	3.32
At5g22500	acyl CoA reductase, putative/male-sterility protein, putative	3.32
At4g16590	glucosyltransferase-related	3.28
At2g36570	leucine-rich repeat transmembrane protein kinase, putative	3.28
At3g44750	histone deacetylase, putative (HD2A)	3.28
At2g43510	trypsin inhibitor, putative	3.27
At1g10780	F-box family protein	3.27
At3g18960	transcriptional factor B3 family protein	3.27
At1g34245	expressed protein	3.25
At2g27510	ferredoxin, putative	3.25
At3g12870	expressed protein	3.25
At3g10570	cytochrome P450, putative	3.24
At1g56580	expressed protein	3.24
At5g52120	F-box family protein/SKP1 interacting partner 3-related	3.23
At2g23560	hydrolase, alpha/beta fold family protein	3.23
At5g62080	protease inhibitor/seed storage/lipid transfer protein (LTP) family protein	3.21
At1g14120	2-oxoglutarate-dependent dioxygenase, putative	3.20
At3g51590	lipid transfer protein, putative	3.20
At4g36180	leucine-rich repeat family protein	3.20
At5g62550	expressed protein	3.19
At4g29610	cytidine deaminase, putative/cytidine aminohydrolase, putative	3.19
At2g32280	expressed protein	3.19
At1g17560	ribosomal protein L14 family protein	3.16
At2g21660	glycine-rich RNA-binding protein (GRP7)	3.15
At5g65280	lanthionine synthetase C-like family protein	3.14
At1g72670	calmodulin-binding family protein	3.14
At1g14250	nucleoside phosphatase family protein/GDA1/CD39 family protein	3.14
At2g06925	phospholipase A2 family protein	3.13
At1g46264	heat shock transcription factor family protein	3.11
At1g53035	expressed protein	3.11
At5g25475	expressed protein	3.11
At4g19400	expressed protein	3.11
At1g66760	MATE efflux family protein	3.10
At3g22760	CXC domain containing TSO1-like protein 1 (SOL1)	3.10
At5g51720	expressed protein	3.10
At3g11480	S-adenosyl-L-methionine:carboxyl methyltransferase family protein	3.10
At4g27570	glycosyltransferase family protein	3.09
At4g37490	G2/mitotic-specific cyclin (CYC1)	3.09
At3g22790	kinase interacting family protein	3.09
At3g56330	N2,N2-dimethylguanosine tRNA methyltransferase family protein	3.09
At2g34640	expressed protein	3.08
At1g31335	expressed protein	3.08
At4g13410	glycosyl transferase family 2 protein	3.08
At5g02810	pseudo-response regulator 7 (APRR7)	3.07
At2g27970	cyclin-dependent kinase, putative/CDK, putative	3.07
At3g23670	phragmoplast-associated kinesin-related protein, putative	3.07
At2g26330	leucine-rich repeat protein kinase, putative (ERECTA)	3.06
At5g55520	expressed protein	3.06
At5g57590	aminotransferase class III family protein	3.05
At3g44590	60S acidic ribosomal protein P2 (RPP2D)	3.05
At1g14890	invertase/pectin methylesterase inhibitor family protein	3.04
At3g20260	expressed protein	3.03
At2g45490	protein kinase, putative	3.03
At3g05180	GDSL-motif lipase/hydrolase family protein	3.03
At1g60270	pseudogene, glycosyl hydrolase family 1	3.02
At4g25630	fibrillarin 2 (FIB2)	3.02
At1g27030	expressed protein	3.02
At5g55120	expressed protein	3.01
At2g29550	tubulin beta-7 chain (TUB7)	3.01
At1g57820	zinc finger (C3HC4-type RING finger) family protein	3.01
At4g36010	pathogenesis-related thaumatin family protein	3.01
At5g15160	bHLH family protein	3.01
At2g01610	invertase/pectin methylesterase inhibitor family protein	3.00
At5g45950	GDSL-motif lipase/hydrolase family protein	2.99
At5g44050	MATE efflux family protein	2.99
At1g23000	heavy-metal-associated domain-containing protein	2.99
At5g56580	mitogen-activated protein kinase kinase (MAPKK), putative (MKK6)	2.99
At3g05020	acyl carrier protein 1, chloroplast (ACP-1)	2.99
At1g02205	CER1 protein	2.98
At5g48310	expressed protein	2.98
At3g19590	WD-40 repeat family protein/mitotic checkpoint protein, putative	2.95
At5g55730	fasciclin-like arabinogalactan-protein (FLA1)	2.95
At1g13650	expressed protein	2.94
At1g56300	DNAJ heat shock N-terminal domain-containing protein	2.94
At1g02800	endo-1,4-beta-glucanase/cellulase (CEL2)	2.94
At1g45130	beta-galactosidase, putative/lactase, putative	2.94
At5g46690	basic helix-loop-helix (bHLH) family protein	2.93
At2g47180	galactinol synthase, putative	2.93
At4g10270	wound-responsive family protein	2.93
At2g39900	LIM domain-containing protein	2.93
At4g26760	microtubule associated protein (MAP65/ASE1) family protein	2.92
At1g78430	tropomyosin-related	2.92
At1g62540	flavin-containing monooxygenase family protein/FMO family protein	2.92
At4g29360	glycosyl hydrolase family 17 protein	2.92
At5g67260	cyclin family protein	2.91
At1g44446	chlorophyll a oxygenase (CAO)/chlorophyll b synthase	2.91
At5g22740	glycosyl transferase family 2 protein	2.90
At2g36200	kinesin motor protein-related	2.89
At3g17820	glutamine synthetase (GS1)	2.89
At1g72250	kinesin motor protein-related	2.89
At2g26760	cyclin, putative	2.89
At5g11410	protein kinase family protein	2.89
At5g10160	beta-hydroxyacyl-ACP dehydratase, putative	2.89
At5g44620	cytochrome P450 family protein	2.88
At2g19670	protein arginine N-methyltransferase, putative	2.88
At3g19820	cell elongation protein/DWARF1/DIMINUTO (DIM)	2.88
At1g73330	protease inhibitor, putative (DR4)	2.88
At5g51600	microtubule associated protein (MAP65/ASE1) family protein	2.87
At5g18790	ribosomal protein L33 family protein	2.87
At5g09980	expressed protein	2.86
At4g39950	cytochrome P450 79B2, putative (CYP79B2)	2.86
At4g28250	beta-expansin, putative (EXPB3)	2.85
At3g55660	expressed protein	2.85
At3g04630	expressed protein	2.85
At2g17740	DC1 domain-containing protein	2.85
At4g31805	WRKY family transcription factor	2.84
At4g28160	hydroxyproline-rich glycoprotein family protein	2.84
At2g19970	pathogenesis-related protein, putative	2.84
At4g32980	homeobox protein (ATH1)	2.84
At2g23150	NRAMP metal ion transporter 3 (NRAMP3)	2.83
At3g28220	meprin and TRAF homology domain-containing protein/MATH domain-	2.82
	containing protein
At2g41650	expressed protein	2.82
At1g44350	IAA-amino acid hydrolase 6, putative (ILL6)	2.81
At2g47630	esterase/lipase/thioesterase family protein	2.81
At3g21950	S-adenosyl-L-methionine:carboxyl methyltransferase family protein	2.81
At4g15440	hydroperoxide lyase (HPL1)	2.80
At3g61250	myb family transcription factor (MYB17)	2.80
At1g52000	jacalin lectin family protein	2.80
At4g24175	expressed protein	2.80
At3g13980	expressed protein	2.79
At5g23530	expressed protein	2.79
At5g24160	squalene monooxygenase 1,2/squalene epoxidase 1,2 (SQP1,2)	2.78
At3g07320	glycosyl hydrolase family 17 protein	2.77
At3g51720	expressed protein	2.77
At1g76310	cyclin, putative	2.77
At4g24610	expressed protein	2.76
At3g05980	expressed protein	2.76
At5g48480	expressed protein	2.75
At3g12700	aspartyl protease family protein	2.75
At2g44040	dihydrodipicolinate reductase family protein /// dihydrodipicolinate	2.74
	reductase family protein
At2g43535	trypsin inhibitor, putative	2.73
At1g06180	myb family transcription factor	2.73
At4g27260	auxin-responsive GH3 family protein	2.72
At1g18650	glycosyl hydrolase family protein 17	2.72
At1g72040	deoxynucleoside kinase family	2.72
At4g33400	dem protein-related/defective embryo and meristems protein-related	2.71
At5g02550	expressed protein	2.71
At4g11290	peroxidase, putative	2.71
At3g15680	zinc finger (Ran-binding) family protein	2.71
At2g02990	ribonuclease 1 (RNS1)	2.71
At1g52400	glycosyl hydrolase family 1 protein/beta-glucosidase, putative (BG1)	2.71
At2g16270	expressed protein	2.71
At5g65640	basic helix-loop-helix (bHLH) family protein	2.71
At4g35060	heavy-metal-associated domain-containing protein/copper chaperone	2.71
	(CCH)-related
At1g06040	zinc finger (B-box type) family protein/salt-tolerance protein (STO)	2.70
At1g19850	transcription factor MONOPTEROS (MP)/auxin-responsive protein	2.70
	(IAA24)/auxin response factor 5 (ARF5)
At1g65900	expressed protein	2.70
At3g05130	expressed protein	2.69
At1g17420	lipoxygenase, putative	2.69
At5g15960	stress-responsive protein (KIN1)	2.69
At4g03070	2-oxoglutarate-dependent dioxygenase (AOP1.2)	2.69
At1g59540	kinesin motor protein-related	2.69
At1g67830	GDSL-motif lipase/hydrolase family protein	2.69
At3g49160	pyruvate kinase family protein	2.68
At5g24470	pseudo-response regulator 5 (APRR5)	2.68
At5g39520	expressed protein	2.68
At1g44760	universal stress protein (USP) family protein	2.67
At5g60880	expressed protein	2.67
At3g54090	pfkB-type carbohydrate kinase family protein	2.67
At3g16690	nodulin MtN3 family protein	2.66
At3g11210	GDSL-motif lipase/hydrolase family protein	2.66
At2g42800	leucine-rich repeat family protein	2.66
At3g10310	kinesin motor protein-related	2.66
At5g65410	zinc finger homeobox family protein/ZF-HD homeobox family protein	2.66
At5g06980	expressed protein	2.66
At2g38620	cell divsion control protein, putative	2.65
At1g80270	DNA-binding protein, putative	2.65
At3g57600	AP2 domain-containing transcription factor, putative	2.65
At2g27840	histone deacetylase-related/HD-related	2.65
At4g35850	pentatricopeptide (PPR) repeat-containing protein	2.65
At1g71050	heavy-metal-associated domain-containing protein/copper chaperone	2.65
	(CCH)-related
At1g65620	LOB domain protein 6/lateral organ boundaries domain protein 6	2.65
	(LBD6)/asymmetric leaves2 (AS2)
At5g50335	expressed protein	2.64
At5g15510	expressed protein	2.64
At1g31550	GDSL-motif lipase, putative	2.64
At1g78600	zinc finger (B-box type) family protein	2.63
At5g61880	signaling molecule-related	2.63
At5g64170	dentin sialophosphoprotein-related	2.63
At3g20240	mitochondrial substrate carrier family protein	2.63
At1g16000	expressed protein	2.62
At3g18850	phospholipid/glycerol acyltransferase family protein	2.62
At2g04570	GDSL-motif lipase/hydrolase family protein	2.61
At2g20940	expressed protein	2.61
At5g54670	kinesin-like protein C (KATC)	2.61
At3g07410	Ras-related GTP-binding family protein	2.61
At2g39920	acid phosphatase class B family protein	2.60
At3g62860	esterase/lipase/thioesterase family protein	2.60
At2g44640	expressed protein	2.60
At4g12600	ribosomal protein L7Ae/L30e/S12e/Gadd45 family protein	2.60
At5g62710	leucine-rich repeat family protein/protein kinase family protein	2.60
At5g01370	expressed protein	2.60
At5g01910	hypothetical protein	2.59
At5g14700	cinnamoyl-CoA reductase-related	2.59
At5g25590	expressed protein	2.59
At5g46280	DNA replication licensing factor, putative	2.59
At5g40330	myb family transcription factor	2.58
At3g57150	dyskerin, putative/nucleolar protein NAP57, putative	2.57
At1g61890	MATE efflux family protein	2.57
At5g14920	gibberellin-regulated family protein	2.57
At2g42110	expressed protein	2.57
At1g14620	expressed protein	2.57
At1g05440	expressed protein	2.57
At1g29270	expressed protein	2.57
At1g06620	2-oxoglutarate-dependent dioxygenase, putative	2.57
At1g69690	TCP family transcription factor, putative	2.57
At1g07370	proliferating cell nuclear antigen 1 (PCNA1)	2.56
At5g07180	leucine-rich repeat family protein/protein kinase family protein	2.56
At2g16780	WD-40 repeat protein (MSI2)	2.56
At5g66230	expressed protein	2.56
At4g39260	glycine-rich RNA-binding protein 8 (GRP8) (CCR1)	2.56
At1g11820	glycosyl hydrolase family 17 protein	2.56
At4g16140	proline-rich family protein	2.55
At3g13650	disease resistance response protein-related/dirigent protein-related	2.55
At3g43960	cysteine proteinase, putative	2.55
At5g13520	peptidase M1 family protein	2.55
At4g16515	expressed protein	2.54
At3g28900	60S ribosomal protein L34 (RPL34C)	2.54
At4g22120	early-responsive to dehydration protein-related/ERD protein-related	2.54
At1g30820	CTP synthase, putative/UTP--ammonia ligase, putative	2.54
At2g05790	glycosyl hydrolase family 17 protein	2.53
At1g13280	allene oxide cyclase family protein	2.53
At5g11510	myb family transcription factor (MYB3R4)	2.52
At1g02200	CER1 protein	2.52
At4g20430	subtilase family protein	2.51
At1g32640	basic helix-loop-helix (bHLH) protein (RAP-1)	2.51
At1g70210	cyclin delta-1 (CYCD1)	2.51
At1g55020	lipoxygenase (LOX1)	2.51
At2g25210	60S ribosomal protein L39 (RPL39A)	2.51
At4g34200	D-3-phosphoglycerate dehydrogenase, putative/3-PGDH, putative	2.51
At1g71260	expressed protein	2.51
At1g28230	purine permease (PUP1)	2.51
At5g65080	MADS-box family protein	2.50
At5g07990	flavonoid 3′-monooxygenase/flavonoid 3′-hydroxylase (F3′H)/	2.50
	cytochrome P450 75B1 (CYP75B1)/transparent testa 7 protein (TT7)
At5g48490	protease inhibitor/seed storage/lipid transfer protein (LTP) family protein	2.50
At5g07230	protease inhibitor/seed storage/lipid transfer protein (LTP) family protein	2.49
At4g01080	expressed protein	2.49
At3g50280	transferase family protein	2.49
At1g63710	cytochrome P450, putative	2.49
At1g73870	zinc finger (B-box type) family protein	2.48
At5g67150	transferase family protein	2.48
At5g03390	expressed protein	2.48
At1g28530	expressed protein	2.48
At5g63810	beta-galactosidase, putative/lactase, putative	2.48
At3g54400	aspartyl protease family protein	2.48
At3g03630	cysteine synthase, chloroplast, putative/O-acetylserine (thiol)-lyase,	2.48
	putative/O-acetylserine sulfhydrylase, putative
At4g09510	beta-fructofuranosidase, putative/invertase, putative/saccharase,	2.48
	putative/beta-fructosidase, putative
At4g28680	tyrosine decarboxylase, putative	2.48
At5g09300	2-oxoisovalerate dehydrogenase, putative/3-methyl-2-oxobutanoate	2.47
	dehydrogenase, putative/branched-chain alpha-keto acid
	dehydrogenase E1 alpha subunit, putative
At2g38400	alanine--glyoxylate aminotransferase, putative/beta-alanine-pyruvate	2.47
	aminotransferase, putative/AGT, putative
At3g05810	expressed protein	2.47
At2g38810	histone H2A, putative	2.47
At2g36870	xyloglucan:xyloglucosyl transferase, putative/xyloglucan	2.47
	endotransglycosylase, putative/endo-xyloglucan transferase, putative
At2g20450	60S ribosomal protein L14 (RPL14A)	2.46
At5g39860	bHLH protein	2.46
At2g36400	expressed protein	2.46
At3g51290	proline-rich family protein	2.46
At5g10830	embryo-abundant protein-related	2.46
At4g35730	expressed protein	2.46
At4g15640	expressed protein	2.46
At2g37690	phosphoribosylaminoimidazole carboxylase family protein/AIR	2.46
	carboxylase family protein
At4g39980	2-dehydro-3-deoxyphosphoheptonate aldolase 1/3-deoxy-D-arabino-	2.46
	heptulosonate 7-phosphate synthase 1/DAHP synthetase 1 (DHS1)
At1g72500	inter-alpha-trypsin inhibitor heavy chain-related	2.46
At5g66440	expressed protein	2.46
At1g68500	expressed protein	2.45
At3g13470	chaperonin, putative	2.45
At4g34290	SWIB complex BAF60b domain-containing protein	2.45
At3g11900	amino acid transporter family protein	2.45
At3g52170	expressed protein	2.45
At4g02060	prolifera protein (PRL)/DNA replication licensing factor Mcm7 (MCM7)	2.45
At1g53140	dynamin family protein	2.45
At1g48630	guanine nucleotide-binding family protein/activated protein kinase C	2.45
	receptor, putative/RACK, putative
At3g05640	protein phosphatase 2C, putative/PP2C, putative	2.44
At1g30530	UDP-glucoronosyl/UDP-glucosyl transferase family protein	2.44
At2g28410	expressed protein	2.44
At3g53370	DNA-binding S1FA family protein	2.44
At4g15140	expressed protein	2.44
At3g61490	glycoside hydrolase family 28 protein/polygalacturonase (pectinase)	2.43
	family protein
At5g46700	senescence-associated protein, putative	2.43
At4g25260	invertase/pectin methylesterase inhibitor family protein	2.43
At2g20670	expressed protein	2.43
At3g17170	ribosomal protein S6 family protein (RFC3)	2.43
At2g37340	splicing factor RSZ33 (RSZ33)	2.43
At5g50370	adenylate kinase, putative	2.43
At3g05890	hydrophobic protein (RCI2B)/low temperature and salt responsive	2.43
	protein (LTI6B)
At5g57180	expressed protein	2.43
At4g15620	integral membrane family protein	2.42
At1g10980	expressed protein	2.42
At1g05620	inosine-uridine preferring nucleoside hydrolase family protein	2.42
At3g14760	expressed protein	2.42
At5g19730	pectinesterase family protein	2.42
At5g27450	mevalonate kinase (MK)	2.42
At1g63100	scarecrow transcription factor family protein	2.41
At2g42740	60S ribosomal protein L11 (RPL11A)	2.41
At4g10390	protein kinase family protein	2.41
At4g39860	expressed protein	2.41
At3g11520	cyclin, putative (CYC2)	2.41
At3g25190	nodulin, putative	2.41
At2g37300	expressed protein	2.41
At3g59650	mitochondrial ribosomal protein L51/S25/CI-B8 family protein	2.41
At4g25150	acid phosphatase, putative	2.41
At5g56500	chaperonin, putative	2.40
At5g19090	heavy-metal-associated domain-containing protein	2.40
At5g28490	expressed protein	2.40
At1g65370	meprin and TRAF homology domain-containing protein/MATH domain-	2.40
	containing protein
At5g12080	mechanosensitive ion channel domain-containing protein/MS ion	2.40
	channel domain-containing protein
At1g31700	copper amine oxidase, putative	2.40
At3g61820	aspartyl protease family protein	2.39
At1g14980	10 kDa chaperonin (CPN10)	2.39
At2g43550	trypsin inhibitor, putative	2.39
At3g27220	kelch repeat-containing protein	2.38
At3g02110	serine carboxypeptidase S10 family protein	2.38
At4g30800	40S ribosomal protein S11 (RPS11B)	2.38
At3g11120	60S ribosomal protein L41 (RPL41C) /// 60S ribosomal protein L41	2.38
	(RPL41C)
At5g12110	elongation factor 1B alpha-subunit 1 (eEF1Balpha1)	2.38
At3g06220	transcriptional factor B3 family protein	2.38
At5g06870	polygalacturonase inhibiting protein 2 (PGIP2)	2.38
At3g61100	expressed protein	2.38
At5g15530	biotin carboxyl carrier protein 2 (BCCP2)	2.38
At1g11080	serine carboxypeptidase S10 family protein	2.37
At4g26780	co-chaperone grpE family protein	2.37
At3g58650	expressed protein	2.37
At5g03230	expressed protein	2.37
At5g51910	TCP family transcription factor, putative	2.37
At4g15490	UDP-glucoronosyl/UDP-glucosyl transferase family protein	2.37
At1g30600	subtilase family protein	2.37
At5g02430	WD-40 repeat family protein	2.37
At3g47500	Dof-type zinc finger domain-containing protein	2.37
At3g16950	dihydrolipoamide dehydrogenase 1, plastidic/lipoamide dehydrogenase	2.36
	1 (PTLPD1)
At4g01730	zinc finger (DHHC type) family protein	2.36
At3g23410	alcohol oxidase-related	2.36
At5g15520	40S ribosomal protein S19 (RPS19B)	2.36
At5g45040	cytochrome c6 (ATC6)	2.35
At1g18330	myb family transcription factor	2.35
At2g46640	hypothetical protein	2.35
At4g10955	lipase class 3 family protein	2.35
At5g15780	pollen Ole e 1 allergen and extensin family protein	2.35
At5g38130	transferase family protein	2.35
At1g06640	2-oxoglutarate-dependent dioxygenase, putative	2.35
At4g18970	GDSL-motif lipase/hydrolase family protein	2.35
At2g05990	enoyl-(acyl-carrier protein) reductase (NADH), chloroplast, putative/	2.34
	NADH-dependent enoyl-ACP reductase, putative
At5g22440	60S ribosomal protein L10A (RPL10aC)	2.34
At3g17120	expressed protein	2.34
At2g43010	phytochrome-interacting factor 4 (PIF4)/basic helix-loop-helix protein 9	2.34
	(bHLH9)/short under red-light 2 (SRL2)
At4g34138	UDP-glucoronosyl/UDP-glucosyl transferase family protein	2.34
At1g23790	expressed protein	2.33
At5g01820	CBL-interacting protein kinase 14 (CIPK14)	2.33
At5g43810	pinhead protein (PINHEAD)/zwille protein (ZWILLE)	2.33
At5g07280	leucine-rich repeat protein kinase, putative/extra sporogenous cells	2.33
	(ESP)
At4g37320	cytochrome P450 family protein	2.33
At1g18710	myb family transcription factor (MYB47)	2.33
At3g44320	nitrilase 3 (NIT3)	2.32
At3g54560	histone H2A.F/Z	2.32
At1g19540	isoflavone reductase, putative	2.32
At3g53380	lectin protein kinase family protein	2.32
At2g36350	protein kinase, putative	2.32
At2g37770	aldo/keto reductase family protein	2.31
At3g07390	auxin-responsive protein/auxin-induced protein (AIR12)	2.31
At4g30660	low temperature and salt responsive protein, putative	2.31
At2g17630	phosphoserine aminotransferase, putative	2.31
At5g39320	UDP-glucose 6-dehydrogenase, putative	2.31
At1g29250	expressed protein	2.31
At3g51450	strictosidine synthase family protein	2.31
At4g37870	phosphoenolpyruvate carboxykinase (ATP), putative/PEP	2.31
	carboxykinase, putative/PEPCK, putative
At3g17160	expressed protein	2.30
At1g72630	expressed protein	2.30
At2g34190	xanthine/uracil permease family protein	2.30
At2g23180	cytochrome P450, putative	2.30
At5g57660	zinc finger (B-box type) family protein	2.30
At2g15090	fatty acid elongase, putative	2.30
At4g34710	arginine decarboxylase 2 (SPE2)	2.30
At1g23410	ubiquitin extension protein, putative/40S ribosomal protein S27A	2.30
	(RPS27aA)
At1g44970	peroxidase, putative	2.29
At1g35290	thioesterase family protein	2.29
At1g75580	auxin-responsive protein, putative	2.29
At1g72140	proton-dependent oligopeptide transport (POT) family protein	2.29
At1g04520	33 kDa secretory protein-related	2.29
At1g68650	expressed protein	2.29
At1g22550	proton-dependent oligopeptide transport (POT) family protein	2.29
At2g04032	metal transporter, putative (ZIP7)	2.29
At1g66850	protease inhibitor/seed storage/lipid transfer protein (LTP) family protein	2.28
At3g48460	GDSL-motif lipase/hydrolase family protein	2.28
At3g10870	hydrolase, alpha/beta fold family protein	2.28
At4g15190	—	2.28
At5g23300	dihydroorotate dehydrogenase, mitochondrial/dihydroorotate oxidase/	2.28
	DHOdehase (PYRD)
At1g64450	proline-rich family protein	2.28
At1g16780	vacuolar-type H+-translocating inorganic pyrophosphatase, putative	2.28
At4g28230	expressed protein	2.27
At1g61610	S-locus lectin protein kinase family protein	2.27
At4g03190	F-box family protein (FBL18)	2.27
At2g26180	calmodulin-binding family protein	2.27
At5g47700	60S acidic ribosomal protein P1 (RPP1C)	2.26
At1g76930	proline-rich extensin-like family protein	2.26
At2g28620	kinesin motor protein-related	2.26
At3g45450	Clp amino terminal domain-containing protein /// Clp amino terminal	2.26
	domain-containing protein
At3g14220	GDSL-motif lipase/hydrolase family protein	2.26
At3g25900	homocysteine S-methyltransferase 1 (HMT-1)	2.26
At2g30040	protein kinase family protein	2.26
At1g62660	beta-fructosidase (BFRUCT3)/beta-fructofuranosidase/invertase,	2.26
	vacuolar
At4g26670	mitochondrial import inner membrane translocase subunit	2.25
	Tim17/Tim22/Tim23 family protein
At4g15210	beta-amylase (BMY1)/1,4-alpha-D-glucan maltohydrolase	2.25
At4g20130	ribulose-1,5 bisphosphate carboxylase/oxygenase large subunit N-	2.25
	methyltransferase-related
At3g10610	40S ribosomal protein S17 (RPS17C)	2.25
At1g04820	tubulin alpha-2/alpha-4 chain (TUA4)	2.24
At4g20930	3-hydroxyisobutyrate dehydrogenase, putative	2.24
At3g23940	dehydratase family	2.24
At1g52720	expressed protein	2.24
At2g36885	expressed protein	2.24
At4g34400	transcriptional factor B3 family protein	2.24
At2g29570	proliferating cell nuclear antigen 2 (PCNA2)	2.24
At4g27010	expressed protein	2.24
At2g41540	NAD-dependent glycerol-3-phosphate dehydrogenase family protein	2.23
At2g38940	phosphate transporter (PT2)	2.23
At2g22900	galactosyl transferase GMA12/MNN10 family protein	2.23
At5g52860	ABC transporter family protein	2.23
At2g45190	axial regulator YABBY1 (YABBY1)/abnormal floral organs protein	2.23
	(AFO)/filamentous flower protein (FIL)
At3g23295	expressed protein	2.23
At3g23290	expressed protein	2.23
At5g62360	invertase/pectin methylesterase inhibitor family protein	2.23
At2g31160	expressed protein	2.22
At5g42900	expressed protein	2.22
At4g13170	60S ribosomal protein L13A (RPL13aC)	2.22
At2g14890	arabinogalactan-protein (AGP9)	2.22
At2g33620	DNA-binding family protein/AT-hook protein 1 (AHP1)	2.22
At2g35120	glycine cleavage system H protein, mitochondrial, putative	2.22
At1g80110	expressed protein	2.22
At4g30650	hydrophobic protein, putative/low temperature and salt responsive	2.21
	protein, putative
At4g16730	terpene synthase/cyclase family protein	2.21
At3g52130	protease inhibitor/seed storage/lipid transfer protein (LTP) family protein	2.21
At1g63240	expressed protein	2.21
At1g74420	xyloglucan fucosyltransferase, putative (FUT3)	2.21
At2g16440	DNA replication licensing factor, putative	2.21
At4g26790	GDSL-motif lipase/hydrolase family protein	2.21
At2g20490	nucleolar RNA-binding Nop10p family protein	2.20
At3g57830	leucine-rich repeat transmembrane protein kinase, putative	2.20
At4g25240	multi-copper oxidase type I family protein	2.20
At3g20015	aspartyl protease family protein	2.20
At5g06550	transcription factor jumonji (jmjC) domain-containing protein	2.20
At5g62690	tubulin beta-2/beta-3 chain (TUB2) /// tubulin beta-2/beta-3 chain (TUB2)	2.20
At2g19780	leucine-rich repeat family protein/extensin family protein	2.20
At5g50580	SUMO activating enzyme, putative /// SUMO activating enzyme, putative	2.20
At5g47330	palmitoyl protein thioesterase family protein	2.20
At2g30500	kinase interacting family protein	2.19
At5g53210	basic helix-loop-helix (bHLH) family protein	2.19
At5g05600	oxidoreductase, 2OG-Fe(II) oxygenase family protein	2.19
At1g49320	BURP domain-containing protein	2.19
At1g65890	acyl-activating enzyme 12 (AAE12)	2.19
At5g23400	disease resistance family protein/LRR family protein	2.19
At5g48820	kip-related protein 3 (KRP3)/cyclin-dependent kinase inhibitor 3 (ICK3)	2.19
At5g11550	expressed protein	2.19
At2g37420	kinesin motor protein-related	2.18
At2g33370	60S ribosomal protein L23 (RPL23B)	2.18
At1g21550	calcium-binding protein, putative	2.18
At5g01075	expressed protein	2.18
At1g53070	legume lectin family protein	2.18
At5g15120	expressed protein	2.18
At2g37790	aldo/keto reductase family protein	2.18
At5g54970	expressed protein	2.18
At3g29230	pentatricopeptide (PPR) repeat-containing protein	2.18
At3g18510	expressed protein	2.18
At5g64860	4-alpha-glucanotransferase, putative/disproportionating enzyme,	2.18
	putative
At2g29730	UDP-glucoronosyl/UDP-glucosyl transferase family protein /// UDP-	2.18
	glucoronosyl/UDP-glucosyl transferase family protein
At1g33811	GDSL-motif lipase/hydrolase family protein	2.18
At1g44835	YbaK/prolyl-tRNA synthetase family protein	2.17
At4g17190	farnesyl pyrophosphate synthetase 2 (FPS2)/FPP synthetase 2	2.17
At5g23740	40S ribosomal protein S11 (RPS11C)	2.17
At1g77120	alcohol dehydrogenase (ADH)	2.17
At2g21130	peptidyl-prolyl cis-trans isomerase/cyclophilin (CYP2)/rotamase	2.17
At4g12880	plastocyanin-like domain-containing protein	2.17
At4g15000	60S ribosomal protein L27 (RPL27C)	2.17
At4g05190	kinesin-like protein A, putative	2.16
At3g03130	expressed protein	2.16
At4g12970	expressed protein	2.16
At5g47600	heat shock protein-related	2.16
At5g47455	expressed protein	2.16
At5g26220	ChaC-like family protein	2.16
At3g26290	cytochrome P450 71B26, putative (CYP71B26)	2.16
At4g17240	expressed protein	2.15
At1g23100	10 kDa chaperonin, putative	2.15
At4g04190	expressed protein	2.15
At5g25490	zinc finger (Ran-binding) family protein	2.15
At5g59480	haloacid dehalogenase-like hydrolase family protein	2.15
At1g78260	RNA recognition motif (RRM)-containing protein	2.15
At2g32650	expressed protein /// expressed protein	2.15
At2g06050	12-oxophytodienoate reductase (OPR3)/delayed dehiscence1 (DDE1)	2.15
At1g16530	LOB domain protein 3/lateral organ boundaries domain protein 3	2.15
	(LBD3)
At5g56940	ribosomal protein S16 family protein	2.15
At2g44450	glycosyl hydrolase family 1 protein	2.15
At2g29890	villin 1 (VLN1)	2.14
At3g51400	expressed protein	2.14
At2g37380	expressed protein	2.14
At4g38890	dihydrouridine synthase family protein	2.14
At4g21910	MATE efflux family protein	2.14
At1g65920	regulator of chromosome condensation (RCC1) family protein/zinc	2.14
	finger protein-related
At1g20590	cyclin, putative /// cyclin, putative	2.14
At1g79530	glyceraldehyde 3-phosphate dehydrogenase, cytosolic, putative/NAD-	2.14
	dependent glyceraldehyde-3-phosphate dehydrogenase, putative ///
	glyceraldehyde 3-phosphate dehydrogenase, cytosolic, putative/NAD-
	dependent glyceraldehyde-3-phosphate dehydrogenase, pu
At1g20930	cell division control protein, putative	2.14
At1g35260	Bet v I allergen family protein	2.14
At5g02280	synbindin, putative	2.14
At5g10400	histone H3	2.14
At4g28950	Rac-like GTP-binding protein (ARAC7)	2.14
At4g18280	glycine-rich cell wall protein-related	2.14
At4g23990	cellulose synthase family protein	2.14
At5g08400	expressed protein	2.14
At1g16790	ribosomal protein-related	2.13
At5g65890	ACT domain-containing protein (ACR1)	2.13
At5g54160	quercetin 3-O-methyltransferase 1/flavonol 3-O-methyltransferase 1/	2.13
	caffeic acid/5-hydroxyferulic acid O-methyltransferase (OMT1)
At5g27330	expressed protein	2.13
At5g67280	leucine-rich repeat transmembrane protein kinase, putative	2.13
At5g04620	aminotransferase class I and II family protein	2.13
At1g49760	polyadenylate-binding protein, putative/PABP, putative	2.13
At3g60360	expressed protein	2.13
At4g02150	importin alpha-2 subunit	2.13
At5g26665	pectinacetylesterase, putative	2.13
At3g62700	glutathione-conjugate transporter, putative	2.13
At3g55280	60S ribosomal protein L23A (RPL23aB)	2.12
At3g13160	pentatricopeptide (PPR) repeat-containing protein	2.12
At3g55310	short-chain dehydrogenase/reductase (SDR) family protein	2.12
At5g60930	chromosome-associated kinesin, putative	2.12
At1g58430	GDSL-motif lipase/hydrolase family protein	2.12
At2g13610	ABC transporter family protein	2.11
At1g05210	expressed protein	2.11
At4g13210	pectate lyase family protein	2.11
At2g20515	expressed protein	2.11
At5g61290	flavin-containing monooxygenase family protein/FMO family protein	2.11
At2g38820	expressed protein	2.11
At1g16350	inosine-5′-monophosphate dehydrogenase, putative	2.11
At5g23860	tubulin beta-8 chain (TUB8) (TUBB8)	2.11
At3g04920	40S ribosomal protein S24 (RPS24A)	2.11
At1g49560	myb family transcription factor	2.11
At2g19810	zinc finger (CCCH-type) family protein	2.11
At4g14270	expressed protein	2.11
At3g27360	histone H3	2.10
At2g19720	40S ribosomal protein S15A (RPS15aB)	2.10
At3g13510	expressed protein	2.10
At3g56130	biotin/lipoyl attachment domain-containing protein	2.10
At3g07770	heat shock protein-related	2.10
At1g78170	expressed protein	2.10
At2g40010	60S acidic ribosomal protein P0 (RPP0A)	2.10
At4g17480	palmitoyl protein thioesterase family protein	2.10
At5g43060	cysteine proteinase, putative/thiol protease, putative	2.10
At3g54260	expressed protein	2.10
At5g60670	60S ribosomal protein L12 (RPL12C)	2.10
At3g17840	leucine-rich repeat transmembrane protein kinase, putative	2.09
At2g18400	ribosomal protein L6 family protein	2.09
At1g14180	expressed protein	2.09
At5g63980	3′(2′),5′-bisphosphate nucleotidase/inositol polyphosphate 1-	2.09
	phosphatase/FIERY1 protein (FRY1) (SAL1)
At2g05540	glycine-rich protein	2.09
At2g22240	inositol-3-phosphate synthase isozyme 2/myo-inositol-1-phosphate	2.09
	synthase 2/MI-1-P synthase 2/IPS 2
At5g02940	expressed protein	2.09
At1g72230	plastocyanin-like domain-containing protein	2.09
At3g51090	expressed protein	2.09
At3g28860	multidrug resistance P-glycoprotein, putative	2.09
At2g36380	ABC transporter family protein	2.09
At3g16490	calmodulin-binding family protein	2.09
At4g39200	40S ribosomal protein S25 (RPS25E)	2.09
At2g40360	transducin family protein/WD-40 repeat family protein	2.09
At5g57290	60S acidic ribosomal protein P3 (RPP3B)	2.09
At2g32765	small ubiquitin-like modifier 5 (SUMO)	2.09
At3g14200	DNAJ heat shock N-terminal domain-containing protein	2.08
At5g08610	DEAD box RNA helicase (RH26)	2.08
At3g52580	40S ribosomal protein S14 (RPS14C)	2.08
At3g18080	glycosyl hydrolase family 1 protein	2.08
At3g53740	60S ribosomal protein L36 (RPL36B)	2.08
At4g05390	ferredoxin--NADP(+) reductase, putative/adrenodoxin reductase,	2.08
	putative
At1g73177	expressed protein	2.08
At5g60300	lectin protein kinase family protein	2.08
At2g22610	kinesin motor protein-related	2.08
At2g16630	proline-rich family protein	2.08
At5g57410	expressed protein	2.08
At4g31700	40S ribosomal protein S6 (RPS6A)	2.08
At2g45970	cytochrome P450, putative	2.08
At3g51740	leucine-rich repeat transmembrane protein kinase, putative	2.08
At1g53520	chalcone-flavanone isomerase-related	2.08
At1g51100	expressed protein	2.08
At5g17490	gibberellin response modulator, putative	2.07
At5g57630	CBL-interacting protein kinase 21, putative (CIPK21)	2.07
At4g14080	glycosyl hydrolase family 17 protein/anther-specific protein (A6)	2.07
At4g28590	expressed protein	2.07
At3g09460	expressed protein	2.07
At2g37190	60S ribosomal protein L12 (RPL12A)	2.07
At2g43020	amine oxidase family protein	2.07
At5g27550	kinesin motor protein-related	2.07
At1g02180	ferredoxin-related	2.07
At2g35605	SWIB complex BAF60b domain-containing protein	2.06
At4g26230	60S ribosomal protein L31 (RPL31B)	2.06
At1g50110	branched-chain amino acid aminotransferase 6 (BCAT6)	2.06
At4g05400	expressed protein	2.06
At2g27710	60S acidic ribosomal protein P2 (RPP2B)	2.06
At5g28060	40S ribosomal protein S24 (RPS24B)	2.06
At3g04000	short-chain dehydrogenase/reductase (SDR) family protein	2.06
At1g03360	exonuclease family protein	2.06
At3g49910	60S ribosomal protein L26 (RPL26A)	2.06
At3g02790	zinc finger (C2H2 type) family protein	2.06
At4g10380	major intrinsic family protein/MIP family protein	2.06
At3g12270	protein arginine N-methyltransferase family protein	2.05
At4g01850	S-adenosylmethionine synthetase 2 (SAM2)	2.05
At5g51100	superoxide dismutase (Fe), putative	2.05
At1g64660	Cys/Met metabolism pyridoxal-phosphate-dependent enzyme family	2.05
	protein
At5g62210	embryo-specific protein-related	2.05
At1g29980	expressed protein	2.05
At1g61120	terpene synthase/cyclase family protein	2.05
At3g52550	hypothetical protein	2.05
At2g29440	glutathione S-transferase, putative	2.05
At2g43460	60S ribosomal protein L38 (RPL38A)	2.05
At3g49260	calmodulin-binding family protein	2.05
At1g64170	cation/hydrogen exchanger, putative (CHX16)	2.05
At1g35180	expressed protein /// expressed protein	2.05
At5g24780	vegetative storage protein 2 (VSP2)	2.05
At1g52930	brix domain-containing protein	2.05
At2g03090	expansin, putative (EXP15)	2.05
At2g18990	expressed protein	2.05
At4g29510	protein arginine N-methyltransferase, putative	2.04
At1g01100	60S acidic ribosomal protein P1 (RPP1A)	2.04
At2g24440	expressed protein	2.04
At2g22170	lipid-associated family protein	2.04
At4g34850	chalcone and stilbene synthase family protein	2.04
At5g54530	expressed protein	2.04
At3g44010	40S ribosomal protein S29 (RPS29A)	2.04
At1g19570	dehydroascorbate reductase, putative	2.04
At2g32060	40S ribosomal protein S12 (RPS12C)	2.04
At2g19170	subtilase family protein	2.04
At5g40610	glycerol-3-phosphate dehydrogenase (NAD+)/GPDH	2.04
At3g11670	digalactosyldiacylglycerol synthase 1 (DGD1)/MGDG:MGDG	2.04
	galactosyltransferase/galactolipid galactosyltransferase
At2g22330	cytochrome P450, putative	2.04
At4g34670	40S ribosomal protein S3A (RPS3aB)	2.04
At5g48880	acetyl-CoA C-acyltransferase 1/3-ketoacyl-CoA thiolase 1 (PKT1)	2.04
At3g06840	expressed protein	2.03
At4g26660	expressed protein	2.03
At5g09770	ribosomal protein L17 family protein	2.03
At4g17610	tRNA/rRNA methyltransferase (SpoU) family protein	2.03
At5g61170	40S ribosomal protein S19 (RPS19C)	2.03
At1g18140	laccase family protein/diphenol oxidase family protein	2.03
At1g51610	cation efflux family protein/metal tolerance protein, putative (MTPc4)	2.03
At2g40435	expressed protein	2.02
At3g44970	cytochrome P450 family protein	2.02
At5g59350	expressed protein	2.02
At5g65870	phytosulfokines 5 (PSK5)	2.02
At5g62290	nucleotide-sensitive chloride conductance regulator (ICIn) family protein	2.02
At1g62510	protease inhibitor/seed storage/lipid transfer protein (LTP) family protein	2.02
At3g47750	ABC transporter family protein	2.02
At5g07090	40S ribosomal protein S4 (RPS4B)	2.02
At3g22740	homocysteine S-methyltransferase 3 (HMT-3)	2.01
At3g61840	expressed protein	2.01
At4g37150	esterase, putative	2.01
At4g18910	aquaglyceroporin/NOD26-like major intrinsic protein 2 (NLM2)	2.01
At3g02080	40S ribosomal protein S19 (RPS19A)	2.01
At1g07790	histone H2B, putative	2.01
At2g37390	heavy-metal-associated domain-containing protein	2.01
At1g03110	transducin family protein/WD-40 repeat family protein	2.01
At1g23800	aldehyde dehydrogenase, mitochondrial (ALDH3)	2.01
At2g39250	AP2 domain-containing transcription factor, putative	2.01
At2g25270	expressed protein	2.01
At5g48760	60S ribosomal protein L13A (RPL13aD)	2.01
At3g21740	expressed protein	2.01
At4g33490	nucellin protein, putative	2.01
At3g60490	AP2 domain-containing transcription factor TINY, putative	2.01
At2g32590	barren family protein	2.01
ccb382	—	2.01
At2g41340	eukaryotic rpb5 RNA polymerase subunit family protein	2.01
At2g36170	ubiquitin extension protein 2 (UBQ2)/60S ribosomal protein L40	2.01
	(RPL40A)
At3g54630	expressed protein	2.01
At5g01790	expressed protein	2.01
At1g63160	replication factor C 40 kDa, putative	2.00
At5g27930	protein phosphatase 2C, putative/PP2C, putative	2.00
At1g33940	hypothetical protein	2.00
At5g66750	SNF2 domain-containing protein/helicase domain-containing protein	2.00
At4g29390	40S ribosomal protein S30 (RPS30B)	2.00
At1g79560	FtsH protease, putative	2.00
At3g46560	mitochondrial import inner membrane translocase (TIM9)	2.00
At2g44120	60S ribosomal protein L7 (RPL7C)	2.00
At4g03110	RNA-binding protein, putative	−2.00
At3g02800	tyrosine specific protein phosphatase family protein	−2.00
At2g24280	serine carboxypeptidase S28 family protein	−2.00
At1g04600	myosin, putative	−2.00
At2g45790	eukaryotic phosphomannomutase family protein	−2.01
At1g78200	protein phosphatase 2C, putative	−2.01
At3g24090	glucosamine--fructose-6-phosphate aminotransferase (isomerizing),	−2.01
	putative/hexosephosphate aminotransferase, putative/glucosamine-6-
	phosphate synthase, putative/D-fructose-6-phosphate
	amidotransferase, putative/GLCN6P synthase, putative
At1g31580	expressed protein	−2.01
At3g01440	oxygen evolving enhancer 3 (PsbQ) family protein	−2.01
At4g35240	expressed protein	−2.01
At5g10250	phototropic-responsive protein, putative	−2.01
At1g53870	expressed protein /// expressed protein	−2.01
At1g80180	expressed protein	−2.01
At1g27200	expressed protein	−2.01
At1g06870	signal peptidase, putative /// signal peptidase, putative	−2.01
At5g06320	harpin-induced family protein/HIN1 family protein/harpin-responsive	−2.01
	family protein/NDR1/HIN1-like protein 3
At1g04550	auxin-responsive protein/indoleacetic acid-induced protein 12 (IAA12)	−2.01
At5g06560	expressed protein	−2.02
At3g50810	integral membrane protein, putative	−2.02
At1g50740	expressed protein	−2.02
At5g53870	plastocyanin-like domain-containing protein	−2.02
At3g01080	WRKY family transcription factor	−2.02
At5g49570	transglutaminase-like family protein	−2.02
At1g32060	phosphoribulokinase (PRK)/phosphopentokinase	−2.02
At4g11840	phospholipase D gamma 3/PLD gamma 3 (PLDGAMMA3)	−2.02
At2g41120	expressed protein	−2.02
At5g47070	protein kinase, putative	−2.02
At4g23180	receptor-like protein kinase 4, putative (RLK4)	−2.03
At3g59090	expressed protein	−2.03
At4g09570	calcium-dependent protein kinase, putative CDPK	−2.03
At4g26690	glycerophosphoryl diester phosphodiesterase family protein	−2.03
At1g01790	K+ efflux antiporter, putative (KEA1)	−2.03
At4g26860	alanine racemase family protein	−2.03
At1g69530	expansin, putative (EXP1)	−2.03
At5g07370	inositol polyphosphate 6-/3-15-kinase 2a (IPK2a)	−2.03
At4g35310	calcium-dependent protein kinase, putative/CDPK, putative	−2.03
At1g28380	expressed protein	−2.03
At2g37940	expressed protein	−2.03
At1g64740	tubulin alpha-1 chain (TUA1)	−2.03
At1g76580	SPL1-Related3 protein (SPL1R3)	−2.03
At1g19835	expressed protein	−2.04
At5g55850	nitrate-responsive NOI protein, putative	−2.04
At3g07580	expressed protein	−2.04
At5g13200	GRAM domain-containing protein/ABA-responsive protein-related	−2.04
At3g14620	cytochrome P450, putative	−2.04
At1g72700	haloacid dehalogenase-like hydrolase family protein	−2.04
At2g42360	zinc finger (C3HC4-type RING finger) family protein	−2.04
At3g28200	peroxidase, putative	−2.04
At1g10740	expressed protein	−2.04
At2g40600	appr-1-p processing enzyme family protein	−2.05
At1g11050	protein kinase family protein	−2.05
At4g16860	disease resistance protein (TIR-NBS-LRR class), putative	−2.05
At5g49360	glycosyl hydrolase family 3 protein	−2.05
At1g53430	leucine-rich repeat family protein/protein kinase family protein	−2.05
At5g39080	transferase family protein	−2.05
At3g15760	expressed protein	−2.05
At2g35760	integral membrane family protein	−2.06
At5g11090	serine-rich protein-related	−2.06
At3g05490	rapid alkalinization factor (RALF) family protein	−2.06
At5g14940	proton-dependent oligopeptide transport (POT) family protein	−2.06
At1g32870	no apical meristem (NAM) family protein	−2.06
At3g17410	serine/threonine protein kinase, putative	−2.06
At1g17340	phosphoinositide phosphatase family protein	−2.06
At3g26440	expressed protein	−2.06
At3g13330	expressed protein	−2.06
At4g17080	MORN (Membrane Occupation and Recognition Nexus) repeat-	−2.06
	containing protein/phosphatidylinositol-4-phosphate 5-kinase-related
At1g17360	COP1-interacting protein-related	−2.06
At4g21390	S-locus lectin protein kinase family protein	−2.07
At4g01330	protein kinase family protein	−2.07
At1g61140	SNF2 domain-containing protein/helicase domain-containing protein/	−2.07
	zinc finger protein-related
At5g57760	expressed protein	−2.07
At3g08970	DNAJ heat shock N-terminal domain-containing protein	−2.07
At4g08500	mitogen-activated protein kinase kinase, putative	−2.07
At5g16570	glutamine synthetase, putative	−2.07
At5g66050	expressed protein	−2.07
At4g00490	beta-amylase, putative/1,4-alpha-D-glucan maltohydrolase, putative	−2.07
At3g04220	disease resistance protein (TIR-NBS-LRR class), putative	−2.07
At1g32940	subtilase family protein	−2.08
At1g09970	leucine-rich repeat transmembrane protein kinase, putative	−2.08
At5g16880	VHS domain-containing protein/GAT domain-containing protein	−2.08
At2g40750	WRKY family transcription factor	−2.08
At3g07650	zinc finger (B-box type) family protein	−2.08
At1g19450	integral membrane protein, putative/sugar transporter family protein	−2.09
At5g20110	dynein light chain, putative	−2.09
At1g16880	uridylyltransferase-related	−2.09
At5g04920	vacuolar protein sorting 36 family protein/VPS36 family protein	−2.09
At2g30520	signal transducer of phototropic response (RPT2)	−2.09
At5g18650	zinc finger (C3HC4-type RING finger) family protein	−2.09
At4g35110	expressed protein	−2.09
At3g53950	glyoxal oxidase-related	−2.09
At3g48990	AMP-dependent synthetase and ligase family protein	−2.09
At4g39640	gamma-glutamyltranspeptidase family protein	−2.09
At3g50830	stress-responsive protein, putative	−2.09
At5g46240	inward rectifying potassium channel (KAT1)	−2.09
At1g14370	protein kinase (APK2a)	−2.09
At1g76530	auxin efflux carrier family protein	−2.10
At5g63080	transcription factor jumonji (jmjC) domain-containing protein	−2.10
At5g61600	ethylene-responsive element-binding family protein	−2.10
At2g37950	zinc finger (C3HC4-type RING finger) family protein	−2.10
At4g23130	receptor-like protein kinase 6 (RLK6)	−2.10
At5g42380	calmodulin-related protein, putative	−2.10
At2g17500	auxin efflux carrier family protein	−2.11
At3g08870	lectin protein kinase, putative	−2.11
At1g65190	protein kinase family protein	−2.11
At1g49470	expressed protein	−2.11
At2g30070	potassium transporter (KUP1)	−2.11
At5g52050	MATE efflux protein-related	−2.11
At1g01360	expressed protein	−2.11
At1g18400	basic helix-loop-helix (bHLH) family protein	−2.11
At3g10500	no apical meristem (NAM) family protein	−2.11
At5g17300	myb family transcription factor	−2.11
At3g62410	CP12 domain-containing protein	−2.11
At5g40670	PQ-loop repeat family protein/transmembrane family protein	−2.11
At5g62900	expressed protein	−2.11
At1g58180	carbonic anhydrase family protein	−2.11
At2g16380	SEC14 cytosolic factor family protein/phosphoglyceride transfer family	−2.12
	protein
At1g70000	DNA-binding family protein	−2.12
At5g25140	cytochrome P450 family protein	−2.12
At3g17770	dihydroxyacetone kinase family protein	−2.12
At5g19875	expressed protein	−2.12
At5g03350	legume lectin family protein	−2.12
At2g03710	MADS-box protein (AGL3)	−2.12
At5g26200	mitochondrial substrate carrier family protein	−2.12
At1g69295	beta-1,3-glucanase-related	−2.12
At2g25625	expressed protein	−2.13
At3g60320	expressed protein	−2.13
At2g02420	expressed protein	−2.13
At4g24570	mitochondrial substrate carrier family protein	−2.13
At3g27960	kinesin light chain-related	−2.13
At3g46110	expressed protein	−2.13
At2g16750	protein kinase family protein	−2.13
At3g47510	expressed protein	−2.13
At3g11420	fringe-related protein	−2.13
At4g39140	expressed protein	−2.13
At2g30010	expressed protein	−2.13
At2g23130	arabinogalactan-protein (AGP17)	−2.14
At5g53450	protein kinase family protein	−2.14
At1g64610	WD-40 repeat family protein	−2.14
At5g25280	serine-rich protein-related	−2.14
At4g24700	expressed protein	−2.14
At5g62860	F-box family protein-related	−2.14
At1g44130	nucellin protein, putative	−2.14
At1g27290	expressed protein	−2.14
At2g41090	calmodulin-like calcium-binding protein, 22 kDa (CaBP-22)	−2.14
At1g02170	latex-abundant family protein (AMC1)/caspase family protein	−2.15
At3g60260	phagocytosis and cell motility protein ELMO1-related	−2.15
At1g55850	cellulose synthase family protein	−2.15
At3g51890	expressed protein	−2.15
At5g55560	protein kinase family protein	−2.15
At1g51800	leucine-rich repeat protein kinase, putative	−2.15
At3g54100	expressed protein	−2.15
At5g60280	lectin protein kinase family protein	−2.15
At2g44500	expressed protein	−2.16
At5g35570	expressed protein	−2.16
At5g13190	expressed protein	−2.16
At1g33050	expressed protein	−2.16
At4g27300	S-locus protein kinase, putative	−2.16
At5g36930	disease resistance protein (TIR-NBS-LRR class), putative	−2.16
At1g03370	C2 domain-containing protein/GRAM domain-containing protein	−2.16
At1g03740	protein kinase family protein	−2.16
At2g42580	tetratricopeptide repeat (TPR)-containing protein	−2.16
At2g32860	glycosyl hydrolase family 1 protein	−2.17
At1g67970	heat shock factor protein, putative (HSF5)/heat shock transcription	−2.17
	factor, putative (HSTF5)
At1g22180	SEC14 cytosolic factor family protein/phosphoglyceride transfer family	−2.17
	protein
At1g62840	expressed protein	−2.17
At1g35210	expressed protein	−2.17
At1g17050	geranyl diphosphate synthase, putative/GPPS, putative/	−2.17
	dimethylallyltransferase, putative/prenyl transferase, putative
At4g14220	zinc finger (C3HC4-type RING finger) family protein	−2.17
At2g03730	ACT domain-containing protein (ACR5)	−2.17
At1g77810	galactosyltransferase family protein	−2.17
At5g24810	ABC1 family protein	−2.17
At3g21080	ABC transporter-related	−2.17
At5g21170	5′-AMP-activated protein kinase beta-2 subunit, putative	−2.17
At4g08930	thioredoxin-related	−2.17
At1g56120	leucine-rich repeat family protein/protein kinase family protein	−2.17
At4g33950	protein kinase, putative	−2.17
At5g26770	expressed protein	−2.18
At5g13180	no apical meristem (NAM) family protein	−2.18
At3g07010	pectate lyase family protein	−2.18
At1g75310	DNAJ heat shock N-terminal domain-containing protein	−2.18
At1g10470	two-component responsive regulator/response regulator 4 (ARR4)	−2.18
At3g02170	expressed protein	−2.18
At5g15830	bZIP transcription factor family protein	−2.18
At4g23320	protein kinase family protein	−2.18
At5g45110	ankyrin repeat family protein/BTB/POZ domain-containing protein	−2.18
At1g32530	zinc finger (C3HC4-type RING finger) family protein	−2.18
At1g64330	myosin heavy chain-related	−2.18
At2g01850	xyloglucan:xyloglucosyl transferase/xyloglucan endotransglycosylase/	−2.18
	endo-xyloglucan transferase (EXGT-A3)
At4g17250	expressed protein	−2.19
At5g23510	expressed protein /// expressed protein	−2.19
At3g47090	leucine-rich repeat transmembrane protein kinase, putative	−2.19
At4g22980	expressed protein	−2.19
At1g33110	MATE efflux family protein	−2.19
At3g16830	WD-40 repeat family protein	−2.19
At4g12830	hydrolase, alpha/beta fold family protein	−2.19
At1g59870	ABC transporter family protein	−2.19
At1g15750	WD-40 repeat family protein	−2.19
At1g16500	expressed protein	−2.20
At1g75080	brassinosteroid signalling positive regulator, putative /// brassinosteroid	−2.20
	signalling positive regulator, putative
At5g46500	expressed protein	−2.20
At1g65390	disease resistance protein (TIR class), putative	−2.20
At4g35090	catalase 2	−2.20
At3g46900	copper transporter, putative	−2.20
At1g64360	expressed protein	−2.21
At3g15310	expressed protein	−2.21
At5g03520	Ras-related GTP-binding protein, putative	−2.21
At3g10930	expressed protein	−2.21
At1g63720	expressed protein	−2.21
At1g67920	expressed protein	−2.21
At4g36990	heat shock transcription factor 4 (HSTF4)	−2.21
At3g53670	expressed protein	−2.21
At1g18620	expressed protein	−2.21
At2g17220	protein kinase, putative	−2.21
At4g37610	TAZ zinc finger family protein/BTB/POZ domain-containing protein	−2.21
At2g36430	expressed protein	−2.21
At4g23270	protein kinase family protein	−2.21
At5g62570	calmodulin-binding protein	−2.21
At1g11330	S-locus lectin protein kinase family protein	−2.22
At2g01190	octicosapeptide/Phox/Bem1p (PB1) domain-containing protein	−2.22
At2g39400	hydrolase, alpha/beta fold family protein	−2.22
At4g18010	inositol polyphosphate 5-phosphatase II (IP5PII)	−2.22
At3g17610	bZIP transcription factor family protein/HY5-like protein (HYH)	−2.22
At1g76990	ACT domain containing protein	−2.22
At5g66900	disease resistance protein (CC-NBS-LRR class), putative ve	−2.22
At5g05250	expressed protein	−2.22
At3g13380	leucine-rich repeat family protein/protein kinase family protein	−2.22
At1g79700	ovule development protein, putative	−2.22
At4g00300	fringe-related protein	−2.22
At5g58670	phosphoinositide-specific phospholipase C (PLC1)	−2.22
At1g72790	hydroxyproline-rich glycoprotein family protein	−2.23
At1g33600	leucine-rich repeat family protein	−2.23
At4g08480	mitogen-activated protein kinase, putative	−2.23
At2g45170	autophagy 8e (APG8e)	−2.23
At1g74170	leucine-rich repeat family protein	−2.23
At3g57230	MADS-box protein (AGL16)	−2.23
At3g19010	oxidoreductase, 2OG-Fe(II) oxygenase family protein	−2.23
At2g15830	expressed protein	−2.23
At3g07610	transcription factor jumonji (jmjC) domain-containing protein	−2.23
At5g56140	KH domain-containing protein	−2.23
At5g44070	phytochelatin synthase 1 (PCS1)	−2.23
At2g31070	TCP family transcription factor, putative	−2.23
At2g26600	glycosyl hydrolase family 17 protein	−2.23
At2g34770	fatty acid hydroxylase (FAH1)	−2.24
At1g05575	expressed protein	−2.24
At5g22570	WRKY family transcription factor	−2.24
At3g49790	expressed protein	−2.24
At5g18240	myb family transcription factor	−2.24
At1g34760	14-3-3 protein GF14 omicron (GRF11)	−2.24
At2g40110	yippee family protein	−2.24
At1g16670	protein kinase family protein	−2.24
At2g38210	ethylene-responsive protein, putative	−2.24
At1g21120	O-methyltransferase, putative	−2.24
At5g66890	disease resistance protein (CC-NBS-LRR class), putative	−2.25
At2g22300	ethylene-responsive calmodulin-binding protein, putative (SR1)	−2.25
At3g18290	zinc finger protein-related	−2.25
At4g19960	potassium transporter family protein	−2.25
At3g01350	proton-dependent oligopeptide transport (POT) family protein	−2.25
At3g55470	C2 domain-containing protein	−2.26
At2g29460	glutathione S-transferase, putative	−2.26
At1g29720	protein kinase family protein	−2.26
At5g61560	protein kinase family protein	−2.26
At5g67470	formin homology 2 domain-containing protein/FH2 domain-containing	−2.27
	protein
At5g56750	Ndr family protein	−2.27
At3g15260	protein phosphatase 2C, putative/PP2C, putative	−2.27
At4g21400	protein kinase family protein	−2.27
At5g46490	disease resistance protein (TIR-NBS-LRR class), putative	−2.27
At1g33240	trihelix DNA-binding protein, putative	−2.27
At4g12390	invertase/pectin methylesterase inhibitor family protein	−2.27
At3g04640	glycine-rich protein	−2.28
At2g30550	lipase class 3 family protein	−2.28
At5g46410	NLI interacting factor (NIF) family protein	−2.28
At5g57560	xyloglucan:xyloglucosyl transferase/xyloglucan endotransglycosylase/	−2.28
	endo-xyloglucan transferase (TCH4)
psbB	—	−2.28
At2g46330	arabinogalactan-protein (AGP16)	−2.28
At2g43430	hydroxyacylglutathione hydrolase, mitochondrial/glyoxalase II (GLX2-1)	−2.28
At2g04070	MATE efflux family protein	−2.28
At5g43190	F-box family protein (FBX6)	−2.29
At5g44290	protein kinase family protein	−2.29
At2g41730	expressed protein	−2.29
At1g76700	DNAJ heat shock N-terminal domain-containing protein	−2.29
At2g43820	UDP-glucoronosyl/UDP-glucosyl transferase family protein	−2.29
At4g16000	expressed protein	−2.29
At3g54960	thioredoxin family protein	−2.29
At1g71697	choline kinase, putative	−2.30
At5g54380	protein kinase family protein	−2.30
At2g29720	monooxygenase family protein	−2.30
At5g09290	3′(2′),5′-bisphosphate nucleotidase, putative/inositol polyphosphate 1-	−2.31
	phosphatase, putative
At2g40670	two-component responsive regulator/response regulator 16 (ARR16)	−2.31
At3g57480	zinc finger (C2H2 type, AN1-like) family protein	−2.31
At5g66200	armadillo/beta-catenin repeat family protein	−2.31
At4g29810	mitogen-activated protein kinase kinase (MAPKK) (MKK2)	−2.31
At5g56860	zinc finger (GATA type) family protein	−2.31
At1g32700	zinc-binding family protein	−2.31
At3g47010	glycosyl hydrolase family 3 protein	−2.31
At1g59620	disease resistance protein (CC-NBS class), putative	−2.32
At1g48300	expressed protein	−2.32
At3g59080	aspartyl protease family protein	−2.32
At5g65730	xyloglucan:xyloglucosyl transferase, putative/xyloglucan	−2.32
	endotransglycosylase, putative/endo-xyloglucan transferase, putative
At1g75410	BEL1-like homeodomain 3 protein (BLH3)	−2.32
At1g75170	SEC14 cytosolic factor family protein/phosphoglyceride transfer family	−2.32
	protein
At1g42990	bZIP transcription factor family protein	−2.32
At5g10650	zinc finger (C3HC4-type RING finger) family protein	−2.32
At4g22710	cytochrome P450 family protein	−2.33
At1g21270	wall-associated kinase 2 (WAK2)	−2.33
At4g18880	heat shock transcription factor 21 (HSF21)	−2.33
At2g21185	expressed protein	−2.33
At2g01860	pentatricopeptide (PPR) repeat-containing protein	−2.33
At1g62420	expressed protein	−2.34
At1g05570	callose synthase 1 (CALS1)/1,3-beta-glucan synthase 1	−2.34
At3g45620	transducin family protein/WD-40 repeat family protein	−2.34
At3g16857	two-component responsive regulator family protein/response regulator	−2.35
	family protein
At3g05900	neurofilament protein-related	−2.35
At1g53570	mitogen-activated protein kinase kinase kinase (MAPKKK), putative	−2.35
	(MAP3Ka)
At1g23870	glycosyl transferase family 20 protein/trehalose-phosphatase family	−2.35
	protein
At3g51920	calmodulin-9 (CAM9)	−2.35
At1g80130	expressed protein	−2.35
At3g04030	myb family transcription factor	−2.35
At1g51270	vesicle-associated membrane protein, putative/VAMP	−2.35
At1g26960	homeobox-leucine zipper protein, putative/HD-ZIP transcription factor,	−2.35
	putative
At1g03210	phenazine biosynthesis PhzC/PhzF family protein	−2.35
At5g05460	glycosyl hydrolase family 85 protein	−2.36
At1g77760	nitrate reductase 1 (NR1)	−2.36
At1g31350	F-box family protein	−2.36
At1g61180	disease resistance protein (CC-NBS-LRR class), putative	−2.36
At4g28085	expressed protein	−2.36
At2g15040	pseudogene, disease resistance protein-related	−2.36
At1g74330	protein kinase, putative	−2.37
At5g62620	galactosyltransferase family protein	−2.37
At1g23830	expressed protein	−2.37
At5g08240	expressed protein	−2.37
At2g18230	inorganic pyrophosphatase (soluble) (PPA)/pyrophosphate phospho-	−2.38
	hydrolase/PPase
At2g12290	hypothetical protein /// hypothetical protein	−2.38
At1g22510	zinc finger (C3HC4-type RING finger) family protein	−2.38
At5g67420	LOB domain protein 37/lateral organ boundaries domain protein 37	−2.38
	(LBD37)
At4g17350	expressed protein	−2.38
At4g23480	hydroxyproline-rich glycoprotein family protein	−2.38
At1g44100	amino acid permease 5, putative (AAP5)	−2.38
At5g65210	bZIP family transcription factor (TGA1)	−2.38
At1g77900	expressed protein	−2.38
At3g57530	calcium-dependent protein kinase, putative/CDPK, putative	−2.38
At1g75020	phospholipid/glycerol acyltransferase family protein	−2.39
At1g08570	thioredoxin family protein	−2.39
At5g27350	sugar-porter family protein 1 (SFP1)	−2.40
At1g78510	solanesyl diphosphate synthase (SPS)	−2.40
At4g33010	glycine dehydrogenase (decarboxylating), putative/glycine	−2.40
	decarboxylase, putative/glycine cleavage system P-protein, putative
At1g63890	zinc finger (C3HC4-type RING finger) family protein	−2.40
At5g47220	ethylene-responsive element-binding factor 2 (ERF2)	−2.40
At3g28180	glycosyl transferase family 2 protein	−2.40
At1g67810	Fe—S metabolism associated domain-containing protein	−2.41
At5g46270	disease resistance protein (TIR-NBS-LRR class), putative	−2.41
At1g15890	disease resistance protein (CC-NBS-LRR class), putative	−2.41
At4g26060	expressed protein	−2.42
At1g30260	expressed protein	−2.42
At3g63420	heterotrimeric G protein gamma-subunit (AGG1)	−2.42
At4g38260	expressed protein	−2.42
At3g47250	expressed protein	−2.42
At2g24550	expressed protein	−2.43
At2g44240	expressed protein	−2.43
At4g10960	lipase class 3 family protein	−2.43
At1g78860	curculin-like (mannose-binding) lectin family protein	−2.43
At1g67850	expressed protein	−2.43
At4g26090	disease resistance protein RPS2 (CC-NBS-LRR class), putative	−2.44
At5g40890	chloride channel protein (CLC-a)	−2.44
At5g45500	expressed protein	−2.44
At5g50200	expressed protein	−2.44
At4g36520	trichohyalin-related	−2.44
At4g36500	expressed protein	−2.44
At1g74440	expressed protein	−2.44
At3g22910	calcium-transporting ATPase, plasma membrane-type, putative (ACA13)	−2.45
At5g54490	calcium-binding EF-hand protein, putative	−2.45
At1g12580	protein kinase family protein	−2.45
At3g52800	zinc finger (AN1-like) family protein	−2.45
At4g25000	alpha-amylase, putative/1,4-alpha-D-glucan glucanohydrolase, putative	−2.45
At5g37600	glutamine synthetase, putative	−2.46
At4g34480	glycosyl hydrolase family 17 protein	−2.46
At4g33920	protein phosphatase 2C family protein/PP2C family protein	−2.46
At4g35780	protein kinase family protein	−2.46
At1g04040	acid phosphatase class B family protein	−2.46
At2g45910	protein kinase family protein/U-box domain-containing protein	−2.47
At4g30850	expressed protein	−2.47
At2g01180	phosphatidic acid phosphatase family protein/PAP2 family protein	−2.47
At3g06080	expressed protein	−2.47
At1g77990	sulfate transporter	−2.47
At1g49050	aspartyl protease family protein	−2.47
At1g29240	expressed protein	−2.48
At3g48850	mitochondrial phosphate transporter, putative	−2.48
At3g06150	expressed protein	−2.48
At5g65010	asparagine synthetase 2 (ASN2)	−2.48
At3g63380	calcium-transporting ATPase, plasma membrane-type, putative/Ca(2+)-	−2.49
	ATPase, putative (ACA12)
At3g07780	expressed protein	−2.49
At5g59680	leucine-rich repeat protein kinase, putative	−2.50
At5g56870	beta-galactosidase, putative/lactase, putative	−2.50
At1g29660	GDSL-motif lipase/hydrolase family protein	−2.51
At4g16690	esterase/lipase/thioesterase family protein	−2.51
At1g66970	glycerophosphoryl diester phosphodiesterase family protein	−2.51
At1g71880	sucrose transporter/sucrose-proton symporter (SUC1)	−2.51
At1g59590	expressed protein	−2.51
At5g45550	mob1/phocein family protein	−2.51
At2g26080	glycine dehydrogenase (decarboxylating), putative/glycine	−2.52
	decarboxylase, putative/glycine cleavage system P-protein, putative
At5g51630	disease resistance protein (TIR-NBS-LRR class), putative	−2.52
At2g41110	calmodulin-2/3/5 (CAM2) (CAL1)	−2.52
At5g02490	heat shock cognate 70 kDa protein 2 (HSC70-2) (HSP70-2)	−2.52
At1g21130	O-methyltransferase, putative	−2.52
At3g57700	protein kinase, putative	−2.52
At5g59450	scarecrow-like transcription factor 11 (SCL11)	−2.52
At3g56710	sigA-binding protein	−2.53
At1g71330	ABC transporter family protein	−2.53
At5g53970	aminotransferase, putative	−2.53
At4g18740	expressed protein	−2.53
At1g17620	expressed protein	−2.53
At3g28580	AAA-type ATPase family protein	−2.53
At4g32870	expressed protein	−2.53
At4g37790	homeobox-leucine zipper protein 22 (HAT22)/HD-ZIP protein 22	−2.53
At5g56250	expressed protein	−2.54
At1g26270	phosphatidylinositol 3- and 4-kinase family protein	−2.54
At3g56200	amino acid transporter family protein	−2.54
At2g38790	expressed protein	−2.55
At4g00150	scarecrow-like transcription factor 6 (SCL6)	−2.55
At1g55910	metal transporter, putative (ZIP11)	−2.55
At4g27280	calcium-binding EF hand family protein	−2.55
At5g44820	expressed protein	−2.55
At1g72940	disease resistance protein (TIR-NBS class), putative	−2.55
At4g37640	calcium-transporting ATPase 2, plasma membrane-type/Ca(2+)-	−2.55
	ATPase isoform 2 (ACA2)
At2g17550	expressed protein	−2.56
At2g37460	nodulin MtN21 family protein	−2.56
At1g25550	myb family transcription factor	−2.56
At1g63880	disease resistance protein (TIR-NBS-LRR class), putative	−2.56
At2g27830	expressed protein	−2.57
At5g10695	expressed protein	−2.57
At2g23320	WRKY family transcription factor	−2.57
At4g23260	protein kinase family protein	−2.57
At2g06850	xyloglucan:xyloglucosyl transferase/xyloglucan endotransglycosylase/	−2.57
	endo-xyloglucan transferase (EXT) (EXGT-A1)
At1g09560	germin-like protein (GLP4) (GLP5)	−2.57
At2g40270	protein kinase family protein	−2.57
At3g60160	ABC transporter family protein /// ABC transporter family protein	−2.57
At5g61590	AP2 domain-containing transcription factor family protein	−2.58
At2g17440	leucine-rich repeat family protein	−2.58
At3g28510	AAA-type ATPase family protein	−2.58
At5g40850	urophorphyrin III methylase (UPM1)	−2.58
At4g20070	peptidase M20/M25/M40 family protein	−2.58
At1g04200	expressed protein	−2.59
At2g24850	aminotransferase, putative	−2.59
At1g05840	aspartyl protease family protein	−2.59
At4g20780	calcium-binding protein, putative	−2.59
At1g17610	disease resistance protein-related	−2.59
At1g69840	band 7 family protein	−2.60
At3g11280	myb family transcription factor	−2.60
At3g63080	glutathione peroxidase, putative	−2.61
At1g03400	2-oxoglutarate-dependent dioxygenase, putative	−2.61
At1g18210	calcium-binding protein, putative	−2.61
At4g17900	zinc-binding family protein	−2.61
At4g16370	oligopeptide transporter OPT family protein	−2.62
At2g28400	expressed protein	−2.62
At5g56340	zinc finger (C3HC4-type RING finger) family protein	−2.62
At2g19710	expressed protein	−2.62
At3g24503	aldehyde dehydrogenase (ALDH1a)	−2.62
At3g45260	zinc finger (C2H2 type) family protein	−2.62
At3g16220	expressed protein	−2.63
At5g51460	trehalose-6-phosphate phosphatase (TPPA)	−2.63
At2g43850	ankyrin protein kinase, putative (APK1)	−2.63
At4g29050	lectin protein kinase family protein	−2.63
At1g60140	glycosyl transferase family 20 protein/trehalose-phosphatase family	−2.63
	protein
At5g05190	expressed protein	−2.63
At1g79270	expressed protein	−2.63
At2g40100	chlorophyll A-B binding protein (LHCB4.3)	−2.63
At1g61100	disease resistance protein (TIR class), putative	−2.64
At5g44870	disease resistance protein (TIR-NBS-LRR class), putative	−2.64
At1g24530	transducin family protein/WD-40 repeat family protein	−2.64
At1g05000	tyrosine specific protein phosphatase family protein	−2.64
At4g23210	protein kinase family protein	−2.64
At1g23030	armadillo/beta-catenin repeat family protein/U-box domain-containing	−2.64
	protein
At4g08950	phosphate-responsive protein, putative (EXO)	−2.65
At1g71040	multi-copper oxidase type I family protein	−2.65
At3g55950	protein kinase family protein	−2.65
At4g02330	pectinesterase family protein	−2.66
At3g45290	seven transmembrane MLO family protein/MLO-like protein 3 (MLO3)	−2.66
At1g03020	glutaredoxin family protein	−2.66
At3g56400	WRKY family transcription factor	−2.66
At1g51660	mitogen-activated protein kinase kinase (MAPKK), putative (MKK4)	−2.66
At1g03850	glutaredoxin family protein	−2.67
At3g61190	BON1-associated protein 1 (BAP1)	−2.67
At4g01720	WRKY family transcription factor	−2.67
At3g05200	zinc finger (C3HC4-type RING finger) family protein (ATL6)	−2.67
At2g17040	no apical meristem (NAM) family protein	−2.67
At5g04720	disease resistance protein (CC-NBS-LRR class), putative	−2.67
At2g18700	glycosyl transferase family 20 protein/trehalose-phosphatase family	−2.67
	protein
At4g21940	calcium-dependent protein kinase, putative/CDPK, putative	−2.68
At2g31110	expressed protein	−2.68
At5g15730	serine/threonine protein kinase, putative	−2.68
At1g22930	T-complex protein 11	−2.68
At2g33530	serine carboxypeptidase S10 family protein	−2.68
At4g26070	mitogen-activated protein kinase kinase (MAPKK) (MKK1) (MEK1)	−2.68
At3g22930	calmodulin, putative	−2.69
At5g15870	glycosyl hydrolase family 81 protein	−2.69
At2g18300	basic helix-loop-helix (bHLH) family protein	−2.70
At1g22280	protein phosphatase 2C, putative/PP2C, putative	−2.70
At1g15010	expressed protein	−2.71
At5g44130	fasciclin-like arabinogalactan-protein, putative	−2.71
At5g42050	expressed protein	−2.71
At1g18890	calcium-dependent protein kinase 1 (CDPK1)	−2.71
At2g47730	glutathione S-transferase 6 (GST6)	−2.71
At1g27730	zinc finger (C2H2 type) family protein (ZAT10)/salt-tolerance zinc finger	−2.72
	protein (STZ)
At3g15770	expressed protein	−2.72
At1g79380	copine-related	−2.72
At2g44080	expressed protein	−2.72
At2g26980	CBL-interacting protein kinase 3 (CIPK3)	−2.72
At3g13062	expressed protein	−2.72
At4g29950	microtubule-associated protein	−2.72
At5g05960	protease inhibitor/seed storage/lipid transfer protein (LTP) family protein	−2.72
At3g49780	phytosulfokines 3 (PSK3)	−2.73
At2g15320	leucine-rich repeat family protein	−2.73
At3g07980	protein kinase, putative	−2.73
At4g23190	protein kinase family protein	−2.73
At5g11000	expressed protein	−2.74
At5g14120	nodulin family protein	−2.74
At5g37740	C2 domain-containing protein	−2.74
At3g22231	expressed protein	−2.76
At5g20830	sucrose synthase/sucrose-UDP glucosyltransferase (SUS1)	−2.76
At1g16510	auxin-responsive family protein	−2.76
At1g01140	CBL-interacting protein kinase 9 (CIPK9)	−2.76
At1g34420	leucine-rich repeat family protein/protein kinase family protein	−2.76
At5g64410	oligopeptide transporter OPT family protein	−2.76
At1g05630	endonuclease/exonuclease/phosphatase family protein	−2.77
At1g48320	thioesterase family protein	−2.77
At1g68690	pseudogene, protein kinase family	−2.77
At2g28200	zinc finger (C2H2 type) family protein	−2.77
At1g16170	expressed protein	−2.77
At1g20160	subtilase family protein	−2.78
At1g51280	disease resistance protein (TIR class), putative	−2.78
At5g38210	serine/threonine protein kinase family protein	−2.78
At4g28270	zinc finger (C3HC4-type RING finger) family protein	−2.79
At4g17070	expressed protein	−2.79
At1g23140	C2 domain-containing protein	−2.80
At3g50950	disease resistance protein (CC-NBS-LRR class), putative	−2.80
At3g57550	guanylate kinase 2 (GK-2)	−2.80
At4g30110	ATPase E1-E2 type family protein/haloacid dehalogenase-like	−2.80
	hydrolase family protein/heavy-metal-associated domain-containing
	protein
At3g29240	expressed protein	−2.81
At1g22530	SEC14 cytosolic factor family protein/phosphoglyceride transfer family	−2.81
	protein
At4g11850	phospholipase D gamma 1/PLD gamma 1 (PLDGAMMA1)	−2.81
At1g70250	receptor serine/threonine kinase, putative	−2.81
At2g37540	short-chain dehydrogenase/reductase (SDR) family protein	−2.81
At3g56980	basic helix-loop-helix (bHLH) family protein	−2.81
At1g12200	flavin-containing monooxygenase family protein/FMO family protein	−2.82
At3g05650	disease resistance family protein	−2.82
At1g21310	proline-rich extensin-like family protein	−2.82
At1g70940	auxin transport protein, putative (PIN3)	−2.82
At3g56290	expressed protein	−2.83
At5g18490	expressed protein	−2.83
At4g25940	epsin N-terminal homology (ENTH) domain-containing protein	−2.84
At3g47780	ABC transporter family protein	−2.84
At1g15790	expressed protein	−2.84
At5g05440	expressed protein	−2.84
At1g25560	AP2 domain-containing transcription factor, putative	−2.84
At3g23880	F-box family protein	−2.84
At4g33420	peroxidase, putative	−2.84
At5g23210	serine carboxypeptidase S10 family protein	−2.84
At1g17990	12-oxophytodienoate reductase, putative	−2.85
At1g03290	expressed protein	−2.85
At2g33580	protein kinase family protein/peptidoglycan-binding LysM domain-	−2.85
	containing protein
At3g12740	LEM3 (ligand-effect modulator 3) family protein/CDC50 family protein	−2.86
At4g00970	protein kinase family protein	−2.87
At5g39050	transferase family protein	−2.87
At4g17660	protein kinase, putative	−2.88
At1g69270	leucine-rich repeat family protein/protein kinase family protein	−2.88
At1g72430	auxin-responsive protein-related	−2.89
At1g72930	disease resistance protein (TIR-NBS class), putative	−2.89
At3g09830	protein kinase, putative	−2.90
At1g23840	expressed protein	−2.90
At1g22890	expressed protein	−2.91
At2g48030	endonuclease/exonuclease/phosphatase family protein	−2.91
At5g04930	phospholipid-transporting ATPase 1/aminophospholipid flippase 1/	−2.91
	magnesium-ATPase 1 (ALA1)
At1g52200	expressed protein	−2.91
At1g77510	protein disulfide isomerase, putative	−2.91
At3g46130	myb family transcription factor (MYB48)	−2.92
At3g28007	nodulin MtN3 family protein	−2.93
At2g37970	SOUL heme-binding family protein	−2.94
At2g19130	S-locus lectin protein kinase family protein	−2.94
At3g61210	embryo-abundant protein-related	−2.94
At2g29990	pyridine nucleotide-disulphide oxidoreductase family protein	−2.95
At3g21520	expressed protein	−2.95
At3g26230	cytochrome P450 family protein	−2.95
At1g66160	U-box domain-containing protein	−2.95
At4g17090	beta-amylase (CT-BMY)/1,4-alpha-D-glucan maltohydrolase	−2.96
At4g32300	lectin protein kinase family protein	−2.96
At1g72300	leucine-rich repeat transmembrane protein kinase, putative	−2.97
At3g09020	alpha 1,4-glycosyltransferase family protein/glycosyltransferase sugar-	−2.97
	binding DXD motif-containing protein
At4g21830	methionine sulfoxide reductase domain-containing protein/SelR	−2.97
	domain-containing protein
At2g23680	stress-responsive protein, putative	−2.97
At4g33300	disease resistance protein (CC-NBS-LRR class), putative	−2.98
At3g07720	kelch repeat-containing protein	−2.98
At1g05340	expressed protein	−2.98
At2g34510	expressed protein	−2.98
At1g52290	protein kinase family protein	−2.99
At5g20250	raffinose synthase family protein/seed imbibition protein, putative	−3.00
	(din10)
At4g23240	protein kinase family protein	−3.00
At5g46450	disease resistance protein (TIR-NBS-LRR class), putative	−3.00
At1g12290	disease resistance protein (CC-NBS-LRR class), putative	−3.00
At5g60900	lectin protein kinase family protein	−3.01
At1g21000	zinc-binding family protein	−3.01
At5g38900	DSBA oxidoreductase family protein	−3.01
At3g28340	galactinol synthase, putative	−3.02
At3g25600	calmodulin, putative	−3.03
At1g72360	ethylene-responsive element-binding protein, putative	−3.04
At4g35600	protein kinase family protein	−3.04
At1g26420	FAD-binding domain-containing protein	−3.04
At3g09010	protein kinase family protein	−3.04
At5g54860	integral membrane transporter family protein	−3.05
At3g01550	triose phosphate/phosphate translocator, putative	−3.05
At4g13510	ammonium transporter 1, member 1 (AMT1.1)	−3.05
At1g55265	expressed protein	−3.05
At2g04795	expressed protein	−3.05
At3g48640	expressed protein	−3.07
At4g31390	ABC1 family protein	−3.08
At5g25190	ethylene-responsive element-binding protein, putative	−3.08
At5g11970	expressed protein	−3.08
At2g01670	MutT/nudix family protein	−3.09
At4g00330	protein kinase family protein	−3.09
At5g45340	cytochrome P450 family protein	−3.09
At3g26590	MATE efflux family protein	−3.09
At3g19930	sugar transport protein (STP4)	−3.10
At1g34750	protein phosphatase 2C, putative/PP2C, putative	−3.10
At5g46330	leucine-rich repeat transmembrane protein kinase, putative	−3.11
At2g32030	GCN5-related N-acetyltransferase (GNAT) family protein	−3.11
At3g55500	expansin, putative (EXP16)	−3.11
At2g41180	sigA-binding protein-related	−3.12
At4g15530	pyruvate phosphate dikinase family protein	−3.13
At4g22530	embryo-abundant protein-related	−3.13
At1g52870	peroxisomal membrane protein-related	−3.13
At3g07870	F-box family protein	−3.14
At3g62150	multidrug resistant (MDR) ABC transporter, putative	−3.15
At2g29350	tropinone reductase, putative	−3.16
At1g18390	protein kinase family protein	−3.16
At1g67910	expressed protein	−3.17
At4g39070	zinc finger (B-box type) family protein	−3.17
At5g39020	protein kinase family protein	−3.17
At1g35140	phosphate-responsive protein, putative	−3.17
At4g17245	zinc finger (C3HC4-type RING finger) family protein	−3.18
At2g39360	protein kinase family protein	−3.18
At1g68840	DNA-binding protein RAV2 (RAV2)/AP2 domain-containing protein	−3.19
	RAP2.8
At5g57220	cytochrome P450, putative	−3.19
At2g17120	peptidoglycan-binding LysM domain-containing protein	−3.20
At5g45460	expressed protein /// expressed protein	−3.20
At1g68440	expressed protein	−3.20
At5g61210	SNAP25 homologous protein SNAP33 (SNAP33) (SNAP33B)/	−3.21
	synaptosomal-associated protein SNAP25-like 1/snap25a
At5g42760	O-methyltransferase N-terminus domain-containing protein	−3.21
At1g78450	SOUL heme-binding family protein	−3.23
At1g63900	zinc finger (C3HC4-type RING finger) family protein	−3.23
At1g78820	curculin-like (mannose-binding) lectin family protein/PAN domain-	−3.23
	containing protein
At5g67370	expressed protein	−3.23
At1g66960	lupeol synthase, putative/2,3-oxidosqualene-triterpenoid cyclase,	−3.24
	putative
At5g15850	zinc finger protein CONSTANS-LIKE 1 (COL1)	−3.24
At5g03120	expressed protein	−3.24
At5g25350	F-box family protein	−3.25
At1g17600	disease resistance protein (TIR-NBS-LRR class), putative	−3.25
At4g13810	disease resistance family protein/LRR family protein	−3.25
At3g28210	zinc finger (AN1-like) family protein	−3.28
At3g28540	AAA-type ATPase family protein	−3.28
At2g30770	cytochrome P450 71A13, putative (CYP71A13)	−3.28
At4g33790	acyl CoA reductase, putative	−3.28
At5g62165	MADS-box protein (AGL42)	−3.30
At1g30410	ATP-binding cassette transport protein, putative	−3.30
At5g14760	L-aspartate oxidase family protein	−3.30
At1g76960	expressed protein	−3.30
At4g11000	ankyrin repeat family protein	−3.30
At3g11840	U-box domain-containing protein	−3.31
At1g35350	EXS family protein/ERD1/XPR1/SYG1 family protein	−3.31
At2g27920	serine carboxypeptidase S10 family protein	−3.32
At5g02290	protein kinase, putative	−3.33
At1g80840	WRKY family transcription factor	−3.34
At5g49730	ferric reductase-like transmembrane component family protein	−3.36
At1g56150	auxin-responsive family protein	−3.36
At3g55980	zinc finger (CCCH-type) family protein	−3.37
At4g01700	chitinase, putative	−3.37
At3g11820	syntaxin 121 (SYP121)/syntaxin-related protein (SYR1)	−3.37
At1g59710	expressed protein	−3.38
At4g11900	S-locus lectin protein kinase family protein	−3.38
At5g66210	calcium-dependent protein kinase family protein CDPK	−3.38
At4g08850	leucine-rich repeat family protein/protein kinase family protein	−3.40
At5g45000	Toll-Interleukin-Resistance (TIR) domain-containing protein	−3.40
At5g55420	hypothetical protein	−3.41
At1g69720	heme oxygenase 3 (HO3)	−3.41
At4g20830	FAD-binding domain-containing protein	−3.42
At2g22500	mitochondrial substrate carrier family protein	−3.42
At5g44810	expressed protein	−3.43
At5g48380	leucine-rich repeat family protein/protein kinase family protein	−3.43
At3g06070	expressed protein	−3.44
At1g37130	nitrate reductase 2 (NR2)	−3.45
At3g47480	calcium-binding EF hand family protein	−3.45
At4g39800	inositol-3-phosphate synthase isozyme 1/myo-inositol-1-phosphate	−3.45
	synthase 1/MI-1-P synthase 1/IPS 1
At3g60290	oxidoreductase, 2OG-Fe(II) oxygenase family protein	−3.45
At4g16990	disease resistance protein (TIR-NBS class), putative	−3.46
At3g48720	transferase family protein	−3.47
At4g05020	NADH dehydrogenase-related	−3.47
At3g56410	expressed protein	−3.48
At4g35180	amino acid transporter family protein	−3.49
At3g44630	disease resistance protein RPP1-WsB-like (TIR-NBS-LRR class),	−3.49
	putative
At3g07340	basic helix-loop-helix (bHLH) family protein	−3.49
At4g08470	mitogen-activated protein kinase, putative	−3.52
At5g40780	lysine and histidine specific transporter, putative	−3.52
At5g47560	sodium/dicarboxylate cotransporter, putative	−3.53
At3g30180	cytochrome P450, putative	−3.54
psbT	—	−3.54
At2g23810	senescence-associated family protein	−3.54
At3g21870	cyclin family protein	−3.54
At2g32540	cellulose synthase family protein	−3.54
At5g45380	sodium:solute symporter family protein	−3.55
At1g76890	trihelix DNA-binding protein/GT-2 factor (GT2)	−3.56
At1g54740	expressed protein	−3.56
At5g27420	zinc finger (C3HC4-type RING finger) family protein	−3.56
At1g10550	xyloglucan:xyloglucosyl transferase, putative/xyloglucan	−3.57
	endotransglycosylase, putative/endo-xyloglucan transferase, putative
At5g25440	protein kinase family protein	−3.57
At3g44400	disease resistance protein (TIR-NBS-LRR class), putative	−3.57
At1g68570	proton-dependent oligopeptide transport (POT) family protein	−3.58
psbI	—	−3.58
At2g25735	expressed protein	−3.60
At5g24110	WRKY family transcription factor	−3.60
At5g61010	exocyst subunit EXO70 family protein	−3.61
At4g39030	enhanced disease susceptibility 5 (EDS5)/salicylic acid induction	−3.63
	deficient 1 (SID1)
At3g16530	legume lectin family protein	−3.65
At1g55450	embryo-abundant protein-related	−3.66
At1g30250	expressed protein	−3.66
At3g53150	UDP-glucoronosyl/UDP-glucosyl transferase family protein	−3.70
At3g45640	mitogen-activated protein kinase, putative/MAPK, putative (MPK3)	−3.70
At4g39890	Ras-related GTP-binding family protein	−3.71
At3g48090	disease resistance protein (EDS1)	−3.72
At3g60470	hypothetical protein	−3.72
At2g19650	DC1 domain-containing protein	−3.74
At1g71390	disease resistance family protein/LRR family protein	−3.75
At1g24140	matrixin family protein	−3.76
At1g22740	Ras-related protein (RAB7)/AtRab75/small GTP-binding protein,	−3.76
	putative
At5g02760	protein phosphatase 2C family protein/PP2C family protein	−3.78
At5g58770	dehydrodolichyl diphosphate synthase, putative/DEDOL-PP synthase,	−3.78
	putative
At3g04210	disease resistance protein (TIR-NBS class), putative	−3.79
At1g67470	protein kinase family protein	−3.79
At4g00955	expressed protein	−3.79
At1g33260	protein kinase family protein	−3.81
At2g35980	harpin-induced family protein (YLS9)/HIN1 family protein/harpin-	−3.83
	responsive family protein
At4g13340	leucine-rich repeat family protein/extensin family protein	−3.84
At4g38550	expressed protein	−3.85
At5g41790	COP1-interactive protein 1/CIP1	−3.85
At5g47850	protein kinase, putative	−3.85
At1g47400	expressed protein	−3.86
At5g44580	expressed protein	−3.87
At1g02930	glutathione S-transferase, putative	−3.87
At4g17670	senescence-associated protein-related	−3.87
At5g61900	copine BONZAI1 (BON1)	−3.88
At4g34390	extra-large guanine nucleotide binding protein, putative/G-protein,	−3.88
	putative
At4g37560	formamidase, putative/formamide amidohydrolase, putative	−3.88
At1g22400	UDP-glucoronosyl/UDP-glucosyl transferase family protein	−3.88
At5g52740	heavy-metal-associated domain-containing protein	−3.89
At2g24600	ankyrin repeat family protein	−3.90
At5g46050	proton-dependent oligopeptide transport (POT) family protein	−3.90
psbK	—	−3.92
At1g67520	lectin protein kinase family protein	−3.93
At4g14020	rapid alkalinization factor (RALF) family protein	−3.95
At1g72280	endoplasmic reticulum oxidoreductin 1 (ERO1) family protein	−3.95
At5g05300	expressed protein	−3.97
At5g53370	pectinesterase family protein	−3.98
At3g19030	expressed protein	−3.98
At3g12830	auxin-responsive family protein	−3.98
At2g18680	expressed protein	−3.98
At1g13210	haloacid dehalogenase-like hydrolase family protein	−4.00
At2g17290	calcium-dependent protein kinase isoform 6 (CPK6)	−4.00
At4g20110	vacuolar sorting receptor, putative	−4.01
At5g18670	beta-amylase, putative (BMY3)/1,4-alpha-D-glucan maltohydrolase,	−4.03
	putative
At5g14930	leaf senescence-associated protein (SAG101)	−4.04
At5g58310	hydrolase, alpha/beta fold family protein	−4.04
At4g38560	expressed protein	−4.04
At3g49340	cysteine proteinase, putative	−4.04
At4g01010	cyclic nucleotide-regulated ion channel, putative (CNGC13)	−4.06
At1g78000	sulfate transporter (Sultr1;2)	−4.08
At3g44350	no apical meristem (NAM) family protein	−4.10
At2g47130	short-chain dehydrogenase/reductase (SDR) family protein	−4.10
At4g10120	sucrose-phosphate synthase, putative	−4.10
At3g57460	expressed protein	−4.10
At2g32160	expressed protein	−4.11
At2g13810	aminotransferase class I and II family protein	−4.12
At3g25610	haloacid dehalogenase-like hydrolase family protein	−4.12
At3g14470	disease resistance protein (NBS-LRR class), putative	−4.14
At1g56510	disease resistance protein (TIR-NBS-LRR class), putative	−4.15
At1g03870	fasciclin-like arabinogalactan-protein (FLA9)	−4.16
At5g10380	zinc finger (C3HC4-type RING finger) family protein	−4.17
At2g40140	zinc finger (CCCH-type) family protein	−4.20
At3g13950	expressed protein	−4.22
At5g42830	transferase family protein	−4.23
At2g46440	cyclic nucleotide-regulated ion channel/cyclic nucleotide-gated channel	−4.26
	(CNGC3)
At4g08300	nodulin MtN21 family protein	−4.26
At4g14610	disease resistance RPS2 protein	−4.29
At2g30250	WRKY family transcription factor	−4.31
At5g48540	33 kDa secretory protein-related	−4.37
At3g14770	nodulin MtN3 family protein	−4.38
At1g18570	myb family transcription factor (MYB51)	−4.40
At3g19850	phototropic-responsive NPH3 family protein	−4.40
At5g25930	leucine-rich repeat family protein/protein kinase family protein	−4.41
At4g01750	expressed protein	−4.41
At5g58940	protein kinase family protein	−4.42
At1g11260	glucose transporter (STP1)	−4.42
At5g40450	expressed protein	−4.44
At3g30775	proline oxidase, mitochondrial/osmotic stress-responsive proline	−4.45
	dehydrogenase (POX) (PRO1) (ERD5)
At3g02840	immediate-early fungal elicitor family protein	−4.46
At4g16563	aspartyl protease family protein	−4.47
At1g74360	leucine-rich repeat transmembrane protein kinase, putative	−4.48
At5g53550	transporter, putative	−4.48
At4g31000	calmodulin-binding protein	−4.48
At4g28400	protein phosphatase 2C, putative/PP2C, putative	−4.51
At5g19240	expressed protein	−4.53
At2g15080	disease resistance family protein	−4.57
At2g30930	expressed protein	−4.60
At5g17760	AAA-type ATPase family protein	−4.65
At3g09940	monodehydroascorbate reductase, putative	−4.66
At1g15520	ABC transporter family protein	−4.67
At4g34150	C2 domain-containing protein	−4.70
At5g60800	heavy-metal-associated domain-containing protein	−4.74
At1g16420	latex-abundant protein, putative (AMC8)/caspase family protein	−4.74
At3g57520	alkaline alpha galactosidase, putative	−4.74
At1g19380	expressed protein	−4.75
At2g40000	expressed protein	−4.80
At2g29110	glutamate receptor family protein (GLR2.8) (GLUR9)	−4.84
At1g19250	flavin-containing monooxygenase family protein/FMO family protein	−4.86
At1g72240	expressed protein	−4.87
At2g36970	UDP-glucoronosyl/UDP-glucosyl transferase family protein	−4.87
At4g14400	ankyrin repeat family protein	−4.89
At5g44420	plant defensin protein, putative (PDF1.2a)	−4.91
At3g26220	cytochrome P450 family protein	−4.92
At4g02420	lectin protein kinase, putative	−4.92
At1g67800	copine-related	−4.92
At4g37370	cytochrome P450, putative	−4.92
At4g39830	L-ascorbate oxidase, putative	−4.96
At1g26380	FAD-binding domain-containing protein	−4.99
At1g76040	calcium-dependent protein kinase, putative/CDPK, putative	−4.99
At2g29120	glutamate receptor family protein (GLR2.7)	−5.00
At1g21250	wall-associated kinase 1 (WAK1)	−5.02
At1g21910	AP2 domain-containing transcription factor family protein	−5.13
At4g04220	disease resistance family protein	−5.13
At3g13100	ABC transporter family protein	−5.19
At2g26400	acireductone dioxygenase (ARD/ARD′) family protein	−5.20
At4g28490	leucine-rich repeat transmembrane protein kinase, putative	−5.21
At5g43910	pfkB-type carbohydrate kinase family protein	−5.21
At2g32800	protein kinase family protein	−5.23
At1g76970	VHS domain-containing protein/GAT domain-containing protein	−5.23
At3g23550	MATE efflux family protein	−5.24
At2g13790	leucine-rich repeat family protein/protein kinase family protein	−5.31
At5g60950	phytochelatin synthetase-related	−5.33
At5g24210	lipase class 3 family protein	−5.35
At1g13340	expressed protein	−5.40
At5g46230	expressed protein	−5.46
At2g47800	glutathione-conjugate transporter (MRP4)	−5.50
At5g52810	ornithine cyclodeaminase/mu-crystallin family protein	−5.52
At3g22060	receptor protein kinase-related	−5.53
At4g23610	expressed protein	−5.56
At5g58120	disease resistance protein (TIR-NBS-LRR class), putative	−5.56
At4g12720	MutT/nudix family protein	−5.57
At1g72900	disease resistance protein (TIR-NBS class), putative	−5.59
At5g10760	aspartyl protease family protein	−5.60
At1g64380	AP2 domain-containing transcription factor, putative	−5.62
At1g64500	glutaredoxin family protein	−5.62
At4g23200	protein kinase family protein	−5.64
At1g57560	myb family transcription factor (MYB50)	−5.66
At5g67340	armadillo/beta-catenin repeat family protein/U-box domain-containing	−5.69
	protein
At1g66880	serine/threonine protein kinase family protein	−5.73
At1g56060	expressed protein	−5.78
At1g78290	serine/threonine protein kinase, putative	−5.80
At1g51890	leucine-rich repeat protein kinase, putative	−5.82
At2g26190	calmodulin-binding family protein	−5.82
At1g57990	purine permease-related	−5.87
At5g40690	expressed protein	−5.91
At3g52400	syntaxin, putative (SYP122)	−5.96
At4g28390	ADP, ATP carrier protein, mitochondrial, putative/ADP/ATP	−5.99
	translocase, putative
At5g24530	oxidoreductase, 2OG-Fe(II) oxygenase family protein	−6.00
At4g33050	calmodulin-binding family protein	−6.01
At2g15390	xyloglucan fucosyltransferase, putative (FUT4)	−6.01
At5g11920	glycosyl hydrolase family 32 protein	−6.05
At4g23810	WRKY family transcription factor	−6.05
At1g78410	VQ motif-containing protein	−6.06
At2g04430	MutT/nudix family protein	−6.09
At5g22530	expressed protein	−6.10
At1g01340	cyclic nucleotide-regulated ion channel (CNGC10) (ACBK1)	−6.11
At2g46400	WRKY family transcription factor	−6.14
At2g26020	plant defensin-fusion protein, putative (PDF1.2b)	−6.15
At1g07000	exocyst subunit EXO70 family protein	−6.17
At2g02930	glutathione S-transferase, putative	−6.18
At1g24150	formin homology 2 domain-containing protein/FH2 domain-containing	−6.21
	protein
At1g08450	calreticulin 3 (CRT3)	−6.24
At1g72060	expressed protein	−6.28
At3g45970	expansin family protein (EXPL1)	−6.28
At1g65490	expressed protein	−6.31
At1g75750	gibberellin-regulated protein 1 (GASA1)	−6.33
At3g23110	disease resistance family protein	−6.34
At5g54610	ankyrin repeat family protein	−6.42
At4g18250	receptor serine/threonine kinase, putative	−6.54
At5g57550	xyloglucan:xyloglucosyl transferase (XTR3)	−6.54
At3g07520	glutamate receptor family protein (GLR1.4)	−6.62
At3g24900	disease resistance family protein/LRR family protein	−6.62
At4g17030	expansin-related	−6.63
At5g26920	calmodulin-binding protein	−6.66
At1g74710	isochorismate synthase 1 (ICS1)/isochorismate mutase	−6.78
At2g14560	expressed protein	−6.81
At1g08050	zinc finger (C3HC4-type RING finger) family protein	−6.85
At4g04540	protein kinase family protein	−6.86
At3g26210	cytochrome P450 71B23, putative (CYP71B23)	−6.94
At3g48360	speckle-type POZ protein-related	−6.95
At4g25110	latex-abundant family protein (AMC2)/caspase family protein	−7.00
At3g48080	lipase class 3 family protein/disease resistance protein-related	−7.08
At5g01540	lectin protein kinase, putative	−7.14
At2g20145	expressed protein	−7.19
At3g51330	aspartyl protease family protein	−7.22
At3g11340	UDP-glucoronosyl/UDP-glucosyl transferase family protein	−7.31
At1g08930	early-responsive to dehydration stress protein (ERD6)/sugar	−7.32
	transporter family protein
At5g54720	ankyrin repeat family protein	−7.32
At3g54150	embryo-abundant protein-related	−7.33
At1g58420	expressed protein	−7.42
At4g23310	receptor-like protein kinase, putative	−7.43
At2g30140	UDP-glucoronosyl/UDP-glucosyl transferase family protein	−7.46
At1g43910	AAA-type ATPase family protein	−7.71
At1g19020	expressed protein	−7.80
At4g34135	UDP-glucoronosyl/UDP-glucosyl transferase family protein	−7.88
At1g21240	wall-associated kinase, putative	−7.97
At1g66090	disease resistance protein (TIR-NBS class), putative	−8.05
At2g38470	WRKY family transcription factor	−8.37
At1g73805	calmodulin-binding protein	−8.38
At1g65480	flowering locus T protein (FT)	−8.42
At5g55450	protease inhibitor/seed storage/lipid transfer protein (LTP) family protein	−8.50
At4g13900	pseudogene, similar to NL0D	−8.59
At1g13470	expressed protein	−8.67
At1g33720	cytochrome P450, putative	−8.68
At2g41100	touch-responsive protein/calmodulin-related protein 3, touch-induced	−8.85
	(TCH3)
At4g04490	protein kinase family protein	−9.09
At1g73800	calmodulin-binding protein	−9.17
At4g39670	expressed protein	−9.27
At2g24160	pseudogene, leucine rich repeat protein family	−9.37
At5g06530	ABC transporter family protein	−9.72
At3g48650	pseudogene, At14a-related protein	−9.77
At5g22380	no apical meristem (NAM) family protein	−9.81
At3g50480	broad-spectrum mildew resistance RPW8 family protein	−10.43
At2g31880	leucine-rich repeat transmembrane protein kinase, putative	−10.46
At3g52430	phytoalexin-deficient 4 protein (PAD4)	−10.61
At1g10340	ankyrin repeat family protein	−10.89
At1g01560	mitogen-activated protein kinase, putative/MAPK, putative (MPK11)	−11.05
At2g26560	patatin, putative	−11.05
At5g52750	heavy-metal-associated domain-containing protein	−11.16
At3g50930	AAA-type ATPase family protein	−11.18
At2g32680	disease resistance family protein	−11.25
At4g23220	protein kinase family protein	−11.34
At1g49200	zinc finger (C3HC4-type RING finger) family protein	−11.49
At5g41740	disease resistance protein (TIR-NBS-LRR class), putative	−11.62
At2g18690	expressed protein	−11.74
At5g66640	LIM domain-containing protein-related	−12.13
At1g57630	disease resistance protein (TIR class), putative	−12.32
At5g13320	auxin-responsive GH3 family protein	−12.73
At3g01290	band 7 family protein	−12.90
At3g26830	cytochrome P450 71B15, putative (CYP71B15)	−12.92
At3g45860	receptor-like protein kinase, putative	−12.98
At1g35230	arabinogalactan-protein (AGP5)	−13.21
At2g43570	chitinase, putative	−13.28
At3g13610	oxidoreductase, 2OG-Fe(II) oxygenase family protein	−13.35
At3g01830	calmodulin-related protein, putative	−13.50
At3g22600	protease inhibitor/seed storage/lipid transfer protein (LTP) family protein	−13.54
At5g37260	myb family transcription factor	−14.15
At4g14365	zinc finger (C3HC4-type RING finger) family protein/ankyrin repeat	−14.15
	family protein
At1g33960	avirulence-responsive protein/avirulence induced gene (AIG1)	−14.81
At3g11010	disease resistance family protein/LRR family protein	−15.66
At3g57260	glycosyl hydrolase family 17 protein	−15.68
At1g30900	vacuolar sorting receptor, putative	−15.75
At4g03450	ankyrin repeat family protein	−16.58
At1g09080	luminal binding protein 3 (BiP-3) (BP3)	−16.68
At3g25010	disease resistance family protein	−17.67
At4g00700	C2 domain-containing protein	−17.93
At1g35710	leucine-rich repeat transmembrane protein kinase, putative	−18.12
At5g39670	calcium-binding EF hand family protein	−18.46
At4g04500	protein kinase family protein	−18.76
At5g64000	3′(2′),5′-bisphosphate nucleotidase, putative/inositol polyphosphate 1-	−19.35
	phosphatase, putative
At5g25260	expressed protein	−19.56
At3g60420	expressed protein	−20.17
At3g57240	beta-1,3-glucanase (BG3)	−20.23
At2g04450	MutT/nudix family protein	−20.46
At2g18660	expansin family protein (EXPR3)	−20.82
At4g02380	late embryogenesis abundant 3 family protein/LEA3 family protein	−21.23
At4g11890	protein kinase family protein	−22.62
At5g59670	leucine-rich repeat protein kinase, putative	−25.03
At5g18470	curculin-like (mannose-binding) lectin family protein	−28.39
At4g23140	receptor-like protein kinase 5 (RLK5)	−30.43
At4g23150	protein kinase family protein	−34.62
At1g75040	pathogenesis-related protein 5 (PR-5)	−44.51
At5g52760	heavy-metal-associated domain-containing protein	−44.86
At1g14870	expressed protein	−45.29
At4g10500	oxidoreductase, 2OG-Fe(II) oxygenase family protein	−51.31
At2g14610	pathogenesis-related protein 1 (PR-1)	−54.95

TABLE 3

Common genes significantly up or down regulated 2-fold in Arabidopsis
overexpressing (OE) Arabidopsis GLK1 (AtGLK1) and wheat GLK1 (TaGLK1).
Data analysed using FlexArray for normalization and T-Test

		AtGLK1	TaGLK1
		OE Fold	OE Fold
Transcript ID	Gene Title	Change	Change

At1g77960	Phox-bem1 domain protein	42.25	18.56
At3g47340	glutamine-dependent asparagine synthetase 1 (ASN1)	12.29	5.24
At2g39030	GCN5-related N-acetyltransferase (GNAT) family protein	10.94	3.70
At4g23680	major latex protein-related/MLP-related	9.92	8.76
At5g22500	acyl CoA reductase, putative/male-sterility protein, putative	8.42	3.32
At5g45670	GDSL-motif lipase/hydrolase family protein	8.01	24.42
At3g55970	oxidoreductase, 2OG-Fe(II) oxygenase family protein	8.01	7.33
At5g37300	expressed protein	7.39	5.24
At2g37770	aldo/keto reductase family protein	7.37	2.31
At2g20670	expressed protein	6.28	2.43
At1g13650	expressed protein	5.66	2.94
At3g11480	S-adenosyl-L-methionine:carboxyl methyltransferase family	5.60	3.10
	protein
At4g15210	beta-amylase (BMY1)/1,4-alpha-D-glucan maltohydrolase	5.40	2.25
At3g16460	jacalin lectin family protein	5.15	3.91
At1g73260	trypsin and protease inhibitor family protein/Kunitz family	4.95	3.57
At2g23000	serine carboxypeptidase S10 family protein	4.84	6.04
At1g56650	myb family transcription factor (MYB75)	4.67	3.90
At5g48490	protease inhibitor/seed storage/lipid transfer protein (LTP)	4.49	2.50
	family protein
At1g78170	expressed protein	4.47	2.10
At1g14250	nucleoside phosphatase family protein/GDA1/CD39 family	4.37	3.14
At3g15720	glycoside hydrolase family 28 protein/polygalacturonase	4.31	5.67
	(pectinase)
At5g24780	vegetative storage protein 2 (VSP2)	4.20	2.05
At3g04000	short-chain dehydrogenase/reductase (SDR) family protein	3.99	2.06
At5g36910	thionin (THI2.2)	3.99	16.22
At5g42800	dihydroflavonol 4-reductase (dihydrokaempferol 4-reductase)	3.74	12.64
	(DFR)
At4g35060	heavy-metal-associated domain-containing protein/copper	3.73	2.71
	chaperone (CCH)-related
At5g23820	MD-2-related lipid recognition domain-containing protein/ML	3.58	8.06
	domain-containing protein
At1g35290	thioesterase family protein	3.55	2.29
At1g53885	senescence-associated protein-related	3.54	3.60
At2g47180	galactinol synthase, putative	3.49	2.93
At5g59480	haloacid dehalogenase-like hydrolase family protein	3.47	2.15
At2g15090	fatty acid elongase, putative	3.37	2.30
At2g39920	acid phosphatase class B family protein	3.18	2.60
At3g15950	DNA topoisomerase-related	3.11	12.88
At2g34810	FAD-binding domain-containing protein	3.04	3.41
At3g09260	glycosyl hydrolase family 1 protein	2.97	26.10
At2g38870	protease inhibitor, putative	2.96	6.07
At3g28220	meprin and TRAF homology domain-containing protein/	2.92	2.82
	MATH domain protein
At4g12500	protease inhibitor/seed storage/lipid transfer protein (LTP)	2.86	6.13
	family protein
At2g21650	myb family transcription factor	2.81	4.24
At3g28740	cytochrome P450 family protein	2.81	4.76
At1g73870	zinc finger (B-box type) family protein	2.77	2.48
At4g12490	protease inhibitor/seed storage/lipid transfer protein (LTP)	2.77	6.61
	family protein
At5g24420	glucosamine/galactosamine-6-phosphate isomerase-related	2.69	11.34
At2g43510	trypsin inhibitor, putative	2.69	3.27
At1g16530	LOB domain protein 3/lateral organ boundaries domain	2.67	2.15
	protein 3 (LBD3)
At2g22240	inositol-3-phosphate synthase isozyme 2/myo-inositol-1-	2.66	2.09
	phosphate synthase 2/MI-1-P synthase 2/IPS 2
At4g21910	MATE efflux family protein	2.66	2.14
At5g24160	squalene monooxygenase 1,2/ squalene epoxidase 1,2	2.65	2.78
	(SQP1,2)
At1g64200	vacuolar ATP synthase subunit E, putative	2.64	4.68
At5g05600	oxidoreductase, 2OG-Fe(II) oxygenase family protein	2.59	2.19
At4g39510	cytochrome P450 family protein	2.50	4.42
At5g65020	annexin 2 (ANN2)	2.49	3.32
At5g02940	expressed protein	2.45	2.09
At2g02990	ribonuclease 1 (RNS1)	2.45	2.71
At1g66760	MATE efflux family protein	2.44	3.10
At5g17220	glutathione S-transferase, putative	2.44	7.26
At2g39330	jacalin lectin family protein	2.44	4.08
At1g62660	beta-fructosidase (BFRUCT3)/beta-fructofuranosidase/	2.43	2.26
	invertase, vacuolar
At4g17470	palmitoyl protein thioesterase family protein	2.43	9.67
At5g44050	MATE efflux family protein	2.39	2.99
At4g19430	expressed protein	2.39	10.34
At4g22870	leucoanthocyanidin dioxygenase, putative/anthocyanidin	2.36	10.13
	synthase, putative
At4g16730	terpene synthase/cyclase family protein	2.35	2.21
At1g44350	IAA-amino acid hydrolase 6, putative (ILL6)/IAA-Ala	2.34	2.81
	hydrolase, putative
At4g12480	protease inhibitor/seed storage/lipid transfer protein (LTP)	2.30	4.45
	family protein
At1g55020	lipoxygenase (LOX1)	2.28	2.51
At2g46640	hypothetical protein	2.26	2.35
At2g43535	trypsin inhibitor, putative	2.25	2.73
At1g22550	proton-dependent oligopeptide transport (POT) family protein	2.22	2.29
At4g16590	glucosyltransferase-related	2.22	3.28
At3g55110	ABC transporter family protein	2.20	4.53
At5g15960	stress-responsive protein (KIN1)	2.19	2.69
At4g11650	osmotin-like protein (OSM34)	2.17	3.85
At1g56580	expressed protein	2.17	3.24
At4g01080	expressed protein	2.16	2.49
At1g75580	auxin-responsive protein, putative	2.16	2.29
At5g54060	glycosyltransferase family protein	2.13	6.85
At5g06870	polygalacturonase inhibiting protein 2 (PGIP2)	2.10	2.38
At3g11900	amino acid transporter family protein	2.09	2.45
At1g76790	O-methyltransferase family 2 protein	2.05	8.45
At2g36870	xyloglucan:xyloglucosyl transferase, putative	2.02	2.47
At5g54160	quercetin 3-O-methyltransferase 1/flavonol 3-O-	2.02	2.13
	methyltransferase 1/caffeic acid/5-hydroxyferulic acid O-
	methyltransferase (OMT1)
At3g23410	alcohol oxidase-related	2.01	2.36
At3g05180	GDSL-motif lipase/hydrolase family protein	2.01	3.03
At5g65870	phytosulfokines 5 (PSK5)	2.01	2.02
At5g13220	expressed protein	2.00	4.23
At1g24150	formin homology 2 domain-containing protein/FH2 domain-	−2.02	−6.21
	containing protein
At4g27300	S-locus protein kinase, putative	−2.03	−2.16
At5g54610	ankyrin repeat family protein	−2.06	−6.42
At2g36970	UDP-glucoronosyl/UDP-glucosyl transferase family protein	−2.06	−4.87
At3g57260	glycosyl hydrolase family 17 protein	−2.07	−15.68
At2g22500	mitochondrial substrate carrier family protein	−2.07	−3.42
At1g35230	arabinogalactan-protein (AGP5)	−2.07	−13.21
At2g41180	sigA-binding protein-related	−2.07	−3.12
At1g03210	phenazine biosynthesis PhzC/PhzF family protein	−2.08	−2.35
At3g09940	monodehydroascorbate reductase, putative	−2.08	−4.66
At2g24850	aminotransferase, putative	−2.10	−2.59
At5g60950	phytochelatin synthetase-related	−2.11	−5.33
At1g76960	expressed protein	−2.11	−3.30
At4g23130	receptor-like protein kinase 6 (RLK6)	−2.12	−2.10
At2g26020	plant defensin-fusion protein, putative (PDF1.2b)	−2.12	−6.15
At1g56120	leucine-rich repeat family protein/protein kinase family protein	−2.14	−2.17
At1g08050	zinc finger (C3HC4-type RING finger) family protein	−2.15	−6.85
At4g18250	receptor serine/threonine kinase, putative	−2.16	−6.54
At1g01340	cyclic nucleotide-regulated ion channel (CNGC10) (ACBK1)	−2.17	−6.11
At3g05650	disease resistance family protein	−2.17	−2.82
At1g08450	calreticulin 3 (CRT3)	−2.18	−6.24
At1g57630	disease resistance protein (TIR class), putative	−2.18	−12.32
At4g14400	ankyrin repeat family protein	−2.19	−4.89
At3g24900	disease resistance family protein/LRR family protein	−2.21	−6.62
At2g14610	pathogenesis-related protein 1 (PR-1)	−2.22	−54.95
At5g54720	ankyrin repeat family protein	−2.23	−7.32
At3g50480	broad-spectrum mildew resistance RPW8 family protein	−2.23	−10.43
At5g52810	ornithine cyclodeaminase/mu-crystallin family protein	−2.23	−5.52
At5g25260	expressed protein///expressed protein	−2.25	−19.56
At4g20780	calcium-binding protein, putative	−2.25	−2.59
At5g44420	plant defensin protein, putative (PDF1.2a)	−2.26	−4.91
At4g21830	methionine sulfoxide reductase domain-containing protein/	−2.26	−2.97
	SeIR domain-containing protein
At2g20870	cell wall protein precursor, putative	−2.26	−0.06
At4g23200	protein kinase family protein	−2.27	−5.64
At3g26210	cytochrome P450 71B23, putative (CYP71B23)	−2.27	−6.94
At5g39670	calcium-binding EF hand family protein	−2.27	−18.46
At3g60420	expressed protein	−2.27	−20.17
At2g40750	WRKY family transcription factor	−2.28	−2.08
At5g02760	protein phosphatase 2C family protein/PP2C family protein	−2.28	−3.78
At1g10340	ankyrin repeat family protein	−2.31	−10.89
At1g24020	Bet v I allergen family protein	−2.31	−0.10
At3g51330	aspartyl protease family protein	−2.32	−7.22
At5g58940	protein kinase family protein	−2.34	−4.42
At1g35710	leucine-rich repeat transmembrane protein kinase, putative	−2.34	−18.12
At5g45380	sodium:solute symporter family protein	−2.35	−3.55
At4g11850	phospholipase D gamma 1/PLD gamma 1 (PLDGAMMA1)	−2.37	−2.81
At4g00700	C2 domain-containing protein	−2.39	−17.93
At4g34135	UDP-glucoronosyl/UDP-glucosyl transferase family protein	−2.42	−7.88
At1g52200	expressed protein	−2.42	−2.91
At4g23220	protein kinase family protein	−2.46	−11.34
At2g04450	MutT/nudix family protein	−2.47	−20.46
At4g23150	protein kinase family protein	−2.50	−34.62
At1g21250	wall-associated kinase 1 (WAK1)	−2.52	−5.02
At3g09020	alpha 1,4-glycosyltransferase family protein/	−2.54	−2.97
	glycosyltransferase sugar-binding DXD motif-containing
	protein
At2g02930	glutathione S-transferase, putative	−2.55	−6.18
At3g48640	expressed protein	−2.58	−3.07
At4g14365	zinc finger (C3HC4-type RING finger) family protein/ankyrin	−2.61	−14.15
	repeat family protein
At3g01290	band 7 family protein	−2.63	−12.90
At1g30900	vacuolar sorting receptor, putative	−2.64	−15.75
At4g02380	late embryogenesis abundant LEA 3	−2.67	−21.23
At1g13470	expressed protein	−2.71	−8.67
At3g23110	disease resistance family protein	−2.73	−6.34
At5g59670	leucine-rich repeat protein kinase, putative	-2.75	-25.03
At1g75040	pathogenesis-related protein 5 (PR-5)	−2.76	−44.51
At4g01700	chitinase, putative	−2.79	−3.37
At1g77510	protein disulfide isomerase, putative	−2.82	−2.91
At3g25010	disease resistance family protein	−2.90	−17.67
At4g23140	receptor-like protein kinase 5 (RLK5)	−2.95	−30.43
At2g47130	short-chain dehydrogenase/reductase (SDR) family protein	−2.97	−4.10
At4g11890	protein kinase family protein	−3.03	−22.62
At4g10500	oxidoreductase, 2OG-Fe(II) oxygenase family protein	−3.17	−51.31
At5g52760	heavy-metal-associated domain-containing protein	−3.20	−44.86
At3g48080	lipase class 3 family protein/disease resistance protein-	−3.31	−7.08
	related
At5g52740	heavy-metal-associated domain-containing protein	−3.31	−3.89
At5g59680	leucine-rich repeat protein kinase, putative	−3.34	−2.50
At4g17030	expansin-related	−3.35	−6.63
At3g16530	legume lectin family protein	−3.57	−3.65
At4g03450	ankyrin repeat family protein	−3.60	−16.58
At2g26560	patatin, putative	−3.60	−11.05
At3g23550	MATE efflux family protein	−3.61	−5.24
At1g14870	expressed protein	−3.73	−45.29
At1g67810	Fe—S metabolism associated domain-containing protein	−3.77	−2.41
At3g11010	disease resistance family protein/LRR family protein	−3.82	−15.66
At5g55450	protease inhibitor/seed storage/lipid transfer protein (LTP)	−3.90	−8.50
	family protein
At2g14560	expressed protein	−3.96	−6.81
At1g09080	luminal binding protein 3 (BiP-3) (BP3)	−4.12	−16.68
At4g25110	latex-abundant family protein (AMC2)/caspase family protein	−4.21	−7.00
At4g08300	nodulin MtN21 family protein	−4.23	−4.26
At5g18470	curculin-like (mannose-binding) lectin family protein	−4.23	−28.39
At1g29720	protein kinase family protein	−5.41	−2.26
At3g57240	beta-1,3-glucanase (BG3)	−5.50	−20.23

Example 5

Identification of GLK1 Regulated Genes

As can be seen from FIG. 4, and Table 1, one of the most highly accumulated transcripts encode At1g77960 (FIGS. 5A-5B), a hypothetical protein of unknown function. At1s77960 encodes a protein homologous to the octicosapeptide/Phox/Bem1 p (PB1) domain-containing protein At5g09620.1 and D. discoideum GB:EAL72846.1. A PB1 domain containing protein has been observed to be upregulated in response to the pathogen C. higgensianum in Arabidopsis (Narusaka et al, 2004). PB1 domain containing proteins are observed in MEK kinases (Nakamura and Johnson, 2003) and in activation of NADP oxidase in response to oxidative stress (Ito et al, 2001).

Example 6

Isolation of Wheat GLK1 (TaGLK1)

A full length cDNA of wheat GLK1 (Genbank Accession ABL10089) was isolated by screening ESTs generated from a bread wheat cDNA library challenged with the stem rust pathogen. The sequence of rice GLK1 (OSGLK1, accession AAK50393) was used to screen the wheat ESTs. TaLr1168E10R, dbEST Gen Bank accession no. BQ620355 was identified.
The full length wheat GLK1 (TaGLK1, accession ABL10089) was cloned from wheat mRNA using RT-PCR primers derived from the sequence of TaLr1168E10R. HindIII and EcoRV sites were created on the complete coding region of TaGLK1 (Genbank accession: ABL 10089) using the following primers:

	(SEQ ID NO: 44)
	TaGLK1 5′ primer-cgtaagcttggctgtggtgggtag

	(SEQ ID NO: 45)
	TaGLK1 3′ primer-ttgttgtcgatatcgtgctgctgcta.

The complete coding region of TaGLK1 (Genbank accession: ABL10089) was inserted into the bombardment vector pAct-nos (a gift of J. Simmonds, Agriculture and Agri-Food Canada) containing the rice Actin1 promoter and intron (D McElroy, W Zhang, J Cao, and R Wu, 1900 Plant Cell 2: 193-171) to form pAct-TaGLK1 as indicated in FIG. 6.
Transformation into wheat cv. Fielder by particle bombardment is carried out as described by Jordan (Plant Cell Reports 19:1069-1075, 2000).
Examples of wheat plants expressing TaGLK1 is shown in FIG. 8.

Example 7

Transformation of Arabidopsis with Wheat GLK1 (TaGLK1)

TaGLK1 was inserted into the BamHI and KpnI sites of transformation vector pHS723-TaGLK1, introduced into Agrobacterium tumafaciens strain GV3109 pmp90 and used to transform Arabidopsis as describe for AtGLK1 (see Example 2) using the method described in Savitch et al, (2007, Biochem Biophys Res Comm 359: 234-238).
Transformation of Brassica napus with Arabidopsis GLK1 (AtGLK1):
B. napus cv. Westar was transformed with pHS723-AtGLK1 as described in Savitch et al, 2005, Plant Cell Physiol 46: 1525-1539).
Transformation of Maize with AtGLK1:
AtGLK1, driven by the 2× CaMV 35S promoter, was inserted into the HindIII and BamH1 sites of pTF 101.1 (FIG. 13A) to produce the transformation vector pTF101.1-70S-AtGLK1-noster (FIG. 13B). Agrobacterium tumifaciens (strain EHA101) mediated transformation (Frame et al, 2002, Plant Physiol 129: 14-22) was used to introduce pTF101.1-70S-AtGLK1-poster into F2 immature embryos of the maize hybrid genotype Hi II.
Transformation of Rice with AtGLK1:
AtGLK1, driven by the 2× CaMV 35S promoter, was inserted into the HindIII and BamH1 sites of pTF 101 1 (FIG. 13A) to produce the transformation vector pTF101.1-70S-AtGLK1-noster (FIG. 13B). Agrobacterium tumifaciens (strain EHA101) transformed with pTF101.1-70S-AtGLK1 were introduced into mature embryo derived callus tissues of Oryza japonicum cv. Nipponbare (Hiei et al, 1994, The Plant J 6: 271-282; Toki, 1997, Plant Mol Biology Rep 15:16-21). Transformed calli were selected on 2 mg/L Bialaphos.

Soybean Transformation:

AtGLK1, driven by the 2× CaMV 35S promoter, was inserted into the HindIII and BamH1 sites of pTF 101.1 (FIG. 13A) to produce the transformation vector pTF101.1-70S-AtGLK1-noster (FIG. 13B). Agrobacterium tumifaciens (strain EHA 101) transformed with pTF 101.1-70S-AtGLK1 were introduced into explants of the cultivar Williams 82. Plantlets regenerated from cocultivated explants were screened with Liberty herbicide for transformants (Paz et al, 2006, Plant Cell Rep 25: 248-255).

Example 8

Infection of Wheat Leaves with F. graminearum

Individual wheat seedlings (6-8 inch first leaf) grown on soil were removed from the soil and placed horizontally between two glass rods in a transparent chamber. The roots were covered with a moist filter paper connected by a wick to a small beaker in the chamber to prevent drying. The leaves were infected by lightly pressing a glass Pasteur pipette tip on the axial leaf surface and releasing inoculum containing F. graminearum (50,000 spores/ml) on the leaf surface. The chamber was sealed and placed in the dark for 24 h, then returned to an environmental growth chamber set at 25° C., 16 h light period at 200 μEm⁻². The spread of the lesions were compared between different genotypes from days 1-8. Mock inoculation with water alone were also performed. Results are shown in FIG. 9.

Example 9

Infection of Wheat Heads with Fusarium graminearum

Maturing wheat heads at 50% anthesis were spray individually with 5 ml of F. graminearum inoculum containing 50,000 macroconidia spores/ml. The sprayed plants were placed in a Conviron E15 environmental chamber equipped with a fine water mist that is activated at 30 min intervals and set at 25° C., 16 h light at 200 μEm⁻². Browning of individual florets was recorded daily. Examples of wheat heads following treatment are shown in FIG. 10.
Transgenic wheat (Triticum aestivum cv Fielder) over-expressing wheat TaGLK1 (FIG. 8) shows enhanced resistance to the spread of lesions in wheat leaves inoculated with F. graminearum (FIG. 9) and in the development of Fusarium Head Blight (FHB) symptoms in developing wheat heads (FIG. 10)

REFERENCES

Savitch L V, Allard G, Seki M, Robert L S, Tinker N A, Huner N P, Shinozaki K, Singh J. The effect of overexpression of two Brassica CBF/DREB1-like transcription factors on photosynthetic capacity and freezing tolerance in Brassica napus. Plant Cell Physiol. 2005 September; 46(9):1525-39. Epub 2005 Jul. 16.
Ramesh B. Nair, Richard W. Joy I V, Eugen Kurylo, Xiaohong Shi, Joan Schnaider, Raju S. S. Datla, Wilf A. Keller, and Gopalan Selvaraj. Identification of a CYP84 Family of Cytochrome P450-Dependent Mono-Oxygenase Genes in Brassica napus and Perturbation of Their Expression for Engineering Sinapine Reduction in the Seeds. Plant Physiol, August 2000, Vol. 123, pp. 1623-1634.
Clough S J, and Bent A F. Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J. 1998 December; 16(6):735-43.
Makandar R, Essig J S, Schapaugh M A, Trick H N, Shah J, (2006) Genetically engineered resistance to Fusarium Head Blight in wheat by expression of Arabidopsis NPR1. Mol Plant-Microb Inter 19:123-129

All citations are hereby incorporated by reference.
The present invention has been described with regard to one or more embodiments. However, it will be apparent to persons skilled in the art that a number of variations and modifications can be made without departing from the scope of the invention as defined in the claims.

Claims

1. A method of conferring resistance to a pathogen in a plant, the method comprising:

i) introducing a nucleic acid molecule encoding a GLK1 polypeptide or fragment or variant thereof into a plant cell to produce a transgenic plant cell, such that the nucleic acid molecule is positioned for expression in the transgenic plant cell; and

ii) growing a transgenic plant from the transgenic plant cell,

wherein the GLK1 polypeptide or fragment or variant thereof is over-expressed relative to wild type GLK1 at least 2 fold, whereby the 2 fold over-expression is sufficient to confer resistance to the pathogen in the plant.

2. The method of claim 1 wherein the nucleic acid molecule comprises a nucleic acid sequence substantially identical to the nucleic acid sequence set forth in SEQ ID NO: 1.

3. The method of claim 1 wherein the nucleic acid molecule comprises the nucleic acid sequence set forth in SEQ ID NO: 1.

4. The method of claim 1 wherein the nucleic acid molecule hybridizes under conditions of high stringency to the nucleic acid sequence set forth in SEQ ID NO: 1.

5. The method of claim 1 wherein the GLK1 polypeptide comprises an amino acid sequence substantially identical to the amino acid sequence set forth in SEQ ID NO: 2.

6. The method of claim 1 wherein the GLK1 polypeptide is an Arabidopsis polypeptide.

7. The method of claim 1 wherein the GLK1 polypeptide is selected from the group consisting of a wheat GLK1 polypeptide, a maize GLK1 polypeptide, a rice GLK1 polypeptide, and a soy bean GLK1 polypeptide.

8. The method of claim 1 wherein the plant is selected from the group consisting of canola, Brassica spp., maize, tobacco, alfalfa, potato, ginseng, pea, oat, rice, soybean, wheat, barley, sunflower, and cotton.

9. The method of claim 1 wherein the pathogen is selected from the group consisting of Fusarium graminearum, Sclerotinia sclerotium, Puccinia graminis, Puccinia triticinia, Puccinia recondite, and Phytopthora infestans.

10. A genetic construct comprising a regulatory region operably linked to a nucleic acid molecule encoding a GLK1 polypeptide or fragment or variant thereof.

11. The genetic construct of claim 10 wherein the nucleic acid molecule comprises a nucleic acid sequence substantially identical to the nucleic acid sequence set forth in SEQ ID NO: 1.

12. The genetic construct of claim 10 wherein the nucleic acid molecule comprises the nucleic acid sequence set forth in SEQ ID NO: 1.

13. The genetic construct of claim 10 wherein the nucleic acid molecule hybridizes under conditions of high stringency to the nucleic acid sequence set forth in SEQ ID NO: 1.

14. The genetic construct of claim 10 wherein the GLK1 polypeptide is selected from the group consisting of a wheat GLK1 polypeptide, a maize GLK1 polypeptide, a rice GLK1 polypeptide, and a soy bean GLK1 polypeptide.

15. A pathogen resistant plant comprising a GLK1 nucleic acid molecule that encodes a GLK1 polypeptide, wherein expression of the GLK1 produces at least 2 fold greater GLK1 polypeptide relative to a wild-type plant.

16. A vector comprising the genetic construct of claim 10.

17. A host cell, a transgenic plant or a transgenic seed comprising the genetic construct of claim 10.

18. The host cell, transgenic plant, or transgenic seed of claim 17 wherein the host cell, transgenic plant or transgenic seed is selected from the group consisting of canola, Brassica spp., maize, tobacco, alfalfa, potato, ginseng, pea, oat, rice, soybean, wheat, barley, sunflower, and cotton.

19. A method of increasing the nitrogen status in a plant, the method comprising:

ii) growing a transgenic plant from the transgenic plant cell,

20. A plant, or a plant cell with increased nitrogen status, the plant or plant cell comprising a GLK1 nucleic acid molecule that encodes a GLK1 polypeptide, wherein expression of the GLK1 produces at least 2 fold greater GLK1 polypeptide relative to a wild-type plant. the increase in nitrogen status determined by comparing the plant or plant cell with a plant or plant cell that does not express the GLK1 nucleic acid molecule and determining the levels of expression of ASN1, the colour of the leaves, or both, wherein an increase in the expression of ASN1, a darker green leaf, or both, when compared to the plant that does not express GLK1, indicates increased nitrogen status.